Bax-Responsive genes for drug target identification in yeast and fungi

ABSTRACT

The invention describes the use of nucleic acids and polypeptides which are involved in a pathway eventually leading to programmed cell death of yeast or fungi for the preparation of a medicament for treating diseases associated with yeast or fungi or for the treatment of prolifeative disorders or for preventing apoptosis in certain diseases. Methods are provided to identify compounds which selectively modulate the expression or functionality of said polypeptides in the same or a parallel pathway. Also provided are compounds as well as pharmaceutical compositions, medicaments and vaccines. The invention also comprises new nucleic acid sequences, probes and primers derived thereof, expression vectors and host cells transformed with said vectors, polypeptides and antibodies raised against said polypeptides.

FIELD OF THE INVENTION

[0001] The present invention relates to the identification of genes andproteins encoded thereof from yeast and fungi whose expression ismodulated upon programmed cell death and which genes, proteins orfunctional fragments and equivalents thereof may be used as selectivetargets for drugs to treat infections caused by or associated with yeastand fungi or for the treatment of proliferative disorders or for theprevention of apoptosis in certain diseases.

BACKGROUND TO THE INVENTION

[0002] Invasive fungal infections (e.g. Candida spp., Aspergillus spp.,Fusarium spp., Zygomycetes spp.) (Walsh, 1992) have emerged during thepast two decades as important pathogens causing formidable morbidity andmortality in an increasingly diverse and progressively expandingpopulation of immunocompromised patients. Those with the acquired immunedeficiency syndrome (AIDS) constitute the most rapidly growing group ofpatients at risk for life-threatening mycosis. But fungal Infectionshave also increased in frequency in several populations of othersusceptible hosts, including very-low-birth-weight infants, cancerpatients receiving chemotherapy, organ transplant recipients, burnpatients and surgical patients with complications.

[0003] These fungal infections are not limited to humans and othermammals, but are also important in plants where they can cause diseasesor cause the production of unwanted compounds (e.g. Fusarium spp.,Aspergillus spp., Botritis spp., Cladosporium spp.).

[0004] Although recent advances in antifungal chemotherapy have had animpact on these mycoses, expanding populations of immunocompromisedpatients will require newer approaches to antifungal therapy. Thediscovery of novel antifungal agents is thus an essential element of anynew antifungal therapy.

[0005] Classical approaches for identifying antifungal compounds haverelied almost exclusively on inhibition of fungal or yeast growth as anendpoint. Libraries of natural products, semi-synthetic, or syntheticchemicals are screened for their ability to kill or arrest growth of thetarget pathogen or a related nonpathogenic model organism. These testsare cumbersome and provide no information about a compound's mechanismof action. The promising lead compounds that emerge from such screensmust then be tested for possible host-toxicity and detailed mechanism ofaction studies must subsequently be conducted to identify the affectedmolecular target.

[0006] Cells from multicellular organisms can commit suicide in responseto specific signals or injury by an intrinsic program of cell death.Apoptosis is a form of programmed cell death which leads to eliminationof unnecessary or damaged cells. Cells that are either unwanted orpotentially harmful to the organism undergo the apoptotic process andshow events like cell shrinkage, chromatin condensation, cytoplasmiccondensation, digestion of nuclear DNA, loss of mitochondrial membranepotential, plasma membrane blebbing and phagocytosis of the cell debris(Schwartz, et al. 1993). The Bcl-2 family of proteins is centrallyinvolved in the control of the programmed cell death process (PCD).Proteins of this group belong either to the inhibitors of cell death(Bcl-2, Bcl-X_(L)) or to the group of proteins promoting apoptosis (Bax,Bak) (Oltvai and Korsmeyer 1994; Knudson and Korsmeyer 1997; Reed et al.1998). The ability of the Bcl-2 family of proteins to regulate life anddeath of a cell is conserved across evolution. Finding of homologues ofPCD regulatory genes in plants and animals suggests the possibility thatsome functions involved in this process may originally have evolved inunicellular organisms, before a divergent development between the plantand the animal kingdom had happened (Apte et al. 1995).

[0007] Expression of the pro-apoptotic human or mouse Bax protein inSaccharomyces cerevisiae did induce cell death in this budding yeast(Sato et al. 1994; Greenhalf et al. 1996; Zha et al. 1996). It wasinitially described as a process that resembled autophagy withdissolution of the internal organelles and vacuolisation. The apoptoticfeatures characteristic for multicellular eucaryotic cells likemorphological changes In nuclear shape and chromatin condensation, werenot observed in this yeast (Zha et al. 1996). It was therefore suggestedthat Bax-induced cell death in S. cerevisiae is due to the toxicity ofthe Bax protein itself, mediated by a hypothetical pore-formationwithout any involvement of a death program (Muchmore et al. 1996).

[0008] Bax expression in the fission yeast Schizosaccharomyces pombe didin contrast show some of the typical apoptotic changes like DNAfragmentation, chromatin condensation, dissolution of the nuclearenvelope and cytosolic vacuolisation, suggesting the presence of theevolutionary conserved PCD pathway in this unicellular eucaryote (Ink etal. 1997; Jurgensmeier et al. 1997). Since it is very unlikely thatspecies dependent differences in the toxicity of the Bax protein are thereason for this observed difference between the two yeasts, a bona fidecell death pathway may well be present in S. cerevisiae.

[0009] Recent findings of a yeast mutant in the cell division cycle geneCDC48 show a number of morphological and molecular features that areconsidered typical indicators of apoptosis markers in metazoan cells:exposure of phosphatidylserine on the outer leaflet of the cytoplasmicmembrane, DNA breakage as well as chromatin condensation andfragmentation, supporting the existence of a basic PCD machinery in thisunicellular yeast. This theory was supported by the analysis of a wildtype yeast cell expressing the human Bax protein. Comprehensive testsfor morphological markers of apoptosis did show a series of changes,identical to morphological markers defining apoptosis (Ligr, Madeo etal. 1998). Recent findings from the same group (Madeo et al., 1999)implicate oxygen stress as a general regulator of apoptosis in yeast butthe actual mechanism of Bax lethality in S. cerevisiae remains unclear.It is an aim of the present invention to provide new bax sequences forexpression in yeast and fungi and tools for identifying yeast andcandida functions in the pathways leading to programmed cell death.

[0010] It is an aim of the present invention to provide nucleic acids aswell as polypeptides which represent potential molecular targets for theidentification of new compounds which can be used in alleviatingdiseases or conditions associated with yeast or fungal infections.

[0011] It is a further aim of the present invention to provide uses ofthese nucleic acid and polypeptide molecules for treating diseasesassociated with yeast or fungi or for the preparation of (a)medicament(s) for treating said diseases.

[0012] It is also an aim of the invention to provide pharmaceuticalcompositions and vaccines comprising these nucleic acids orpolypeptides.

[0013] It is also an aim of the present invention to provide vectorscomprising these nucleic acids, as well as host cells transfected ortransformed with said vectors.

[0014] It is also an aim of the invention to provide antibodies againstthese polypeptides, which can be used as such, or in a composition as amedicament for treating diseases associated with yeast and fungi.

[0015] It is another aim of the invention to provide methods toselectively identify compounds or polypeptides capable of inhibiting oractivating expression of the polypeptides of the invention or capable ofselectively modulating expression or functionality of such polypeptides.The nucleic acid and polypeptide molecules alternatively can beincorporated into an assay or kit to identify these compounds orpolypeptides.

[0016] It is also an aim of the invention to provide methods forpreventing infection with yeast or fungi.

[0017] It is a further aim of the invention to provide human homologuesfor the nucleic acids and polypeptides of the invention for use intreating proliferative disorders, such as cancer, or for the preventionof apoptosis in certain diseases, or for the preparation of a medicamentfor treating such disorders or diseases.

[0018] All the aims of the present invention have been met by theembodiments as set out below.

SUMMARY OF THE INVENTION

[0019] Since it has been discovered that the mammalian bax gene triggersapoptotic changes in yeast (Ligr et al., 1998), this can be anindication that the molecular pathways eventually leading to programmedcell death may also be partially present in yeast cells and otherunicellular eukaryotes. Identification of genes involved in this processcould be important for the development of new antifungal therapeutics.

[0020] The present inventors overexpressed the Bax protein in thepathogenic yeast Candida albicans and found that this leads to a similarphenotype. However these results could only be received after havingconstructed a new synthetic BAX gene which could be adequately expressedin this pathogenic organism.

[0021] Furthermore, the present inventors identified a range of specificnucleic acids which are involved in the molecular pathways eventuallyleading to programmed cell death. The present inventors were able toidentify via macro array screening a range of genes involved in apathway eventually leading to programmed cell death in the yeastSaccharomyces cerevisiae. Genes which were differentially expressed(analysed using the Pathways™ software) at different time points afterBax expression are envisaged as candidate genes in the presentinvention.

[0022] Additionally, the invention also relates to Candida spp.homologues of the S. cerevisiae candidate genes and their uses instimulating or preventing cell death in yeast and fungi, especiallypathogenic yeast and fungi are herewith envisaged.

[0023] Furthermore, also part of the invention are the human homologuesof these apoptosis-associated S. cerevisiae nucleic acids andpolypeptides and their potential use in treating proliferative disordersin human and other mammals.

DETAILED DESCRIPTION OF THE INVENTION

[0024] The present invention relates to the use of a nucleic acidmolecule encoding a polypeptide which is involved in a pathwayeventually leading to programmed cell death of yeast or fungi and whichnucleic acid sequence is selected from,

[0025] (a) a nucleic acid encoding a protein having an amino acidsequence as represented in any of SEQ ID NOs 18, 20, 22, 24, 26, 28, 30,32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66,68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100,102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128,130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156,158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184,186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212,214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240,242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268,270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296,298, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314,316, 318, 320, 322, 324, 326, 328, 330, 332, 324, 326, 328, 340, 342,344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370,372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396, 398,400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426,428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454,456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478, 480, 482,484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506, 508, 510,512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538,540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566,568, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584,586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612,614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640,642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668,670, 672, 674, 688, 692, 694, 696, 698, 700, 702, 704, 706, 708, 710,712, 714, 716, 718, 720, 722, 724, 726, 728, 730 and 732, or encoding afunctional equivalent, derivative or bioprecursor of said protein,

[0026] (b) a nucleic acid encoding a protein having an amino acidsequence which is more than 70% similar, preferably more than 75% or 80%similar, more preferably more than 85%, 90% or 95% similar and mostpreferably more than 97% similar to any of the amino acid sequences asrepresented by any of SEQ ID NOs 18, 20, 22, 24, 26, 28, 30, 32, 34, 36,38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72,74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106,108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134,136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162,164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190,192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218,220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246,248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274,276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 290, 292,294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320,322, 324, 326, 328, 330, 332, 324, 326, 328, 340, 342, 344, 346, 348,350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376,378, 380, 382, 384, 386, 388, 390, 392, 394, 396, 398, 400, 402, 404,406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432,434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460,462, 464, 466, 468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488,490, 492, 494, 496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516,518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544,546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 560, 562,564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590,592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618,620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646,648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674,688, 692, 694, 696, 698, 700, 702, 704, 706, 708, 710, 712, 714, 716,718, 720, 722, 724, 726, 728, 730 and 732,

[0027] (c) a nucleic acid encoding a protein having an amino acidsequence which is more than 70% identical, preferably more than 75% or80% identical, more preferably more than 85%, 90% or 95% identical andmost preferably more than 97% identical to any of the amino acidsequences as represented by any of SEQ ID NOs 18, 20, 22, 24, 26, 28,30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64,66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100,102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128,130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156,158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184,186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212,214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240,242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268,270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296,298, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314,316, 318, 320, 322, 324, 326, 328, 330, 332, 324, 326, 328, 340, 342,344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370,372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396, 398,400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426,428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454,456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478, 480, 482,484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506, 508, 510,512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538,540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566,568, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584,586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612,614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640,642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668,670, 672, 674, 688, 692, 694, 696, 698, 700, 702, 704, 706, 708, 710,712, 714, 716, 718, 720, 722, 724, 726, 728, 730 and 732,

[0028] (d) a nucleic acid comprising a sequence as represented in any ofSEQ ID NOs 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45,47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81,83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113,115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141,143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169,171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197,199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225,227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253,255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281,283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309,311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337,339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365,367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393,395, 397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421,423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, 449,451, 453, 455, 457, 459, 461, 463, 465, 467, 469, 471, 473, 475, 477,479, 481, 483, 485, 487, 489, 491, 493, 495, 497, 499, 501, 503, 505,507, 509, 511, 513, 515, 517, 519, 521, 523, 525, 527, 529, 531, 533,535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557, 559, 561,563, 565, 567, 569, 571, 573, 575, 577, 579, 581, 583, 585, 587, 589,591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617,619, 621, 623, 625, 627, 629, 631, 633, 635, 637, 639, 641, 643, 645,647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673,687, 691, 693, 695, 697, 699, 701, 703, 705, 707, 709, 711, 713, 715,717, 719, 721, 723, 725, 727, 729 and 731,

[0029] (e) a nucleic acid which is more than 70% identical, preferablymore than 75 or 80% identical, more preferably more than 85%, or 90% or95% identical and most preferably more than 97% identical to any of thenucleic acid sequences as represented by any of SEQ ID NOs 17, 19, 21,23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57,59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93,95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123,125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151,153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179,181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207,209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235,237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263,265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291,293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319,321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347,349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375,377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399, 401, 403,405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431,433, 435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457, 459,461, 463, 465, 467, 469, 471, 473, 475, 477, 479, 481, 483, 485, 487,489, 491, 493, 495, 497, 499, 501, 503, 505, 507, 509, 511, 513, 515,517, 519, 521, 523, 525, 527, 529, 531, 533, 535, 537, 539, 541, 543,545, 547, 549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569, 571,573, 575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599,601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627,629, 631, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655,657, 659, 661, 663, 665, 667, 669, 671, 673, 687, 691, 693, 695, 697,699, 701, 703, 705, 707, 709, 711, 713, 715, 717, 719, 721, 723, 725,727, 729 and 731,

[0030] (f) a nucleic acid encoding a functional fragment of any of thenucleic acids as specified in a) to e); and

[0031] (g) the complement of any of the nucleic acids as specified in a)to f),

[0032] for the preparation of a medicament for treating diseasesassociated with yeast or fungi. Sequence similarity searches wereperformed using the BLAST software package version 2. Identity andsimilarity percentages were calculated using BLOSUM62 as a scoringmatrix. As known in the art, “similarity” between two polypeptides isdetermined by comparing the amino acid sequence and its conserved aminoacid substitutes of one polypeptide to the sequence of a secondpolypeptide. Moreover, also known in the art is “identity” which meansthe degree of sequence relatedness between two polypeptide or twopolynucleotide sequences as determined by the identity of the matchbetween two strings of such sequences. Both identity and similarity canbe readily calculated. While there exist a number of methods to measureidentity and similarity between two polynucleotide or polypeptidesequences, the terms “identity” and “similarity” are well known toskilled artisans (Carillo and Lipton, 1988). Methods commonly employedto determine identity or similarity between two sequences include, butare not limited to, those disclosed in “Guide to Huge Computers (Bishop,1994) and Carillo and Lipton (1988). Preferred methods to determineidentity are designed to give the largest match between the twosequences tested. Methods to determine identity and similarity arecodified in computer programs. Preferred computer program methods todetermine identity and similarity between two sequences include, but arenot limited to, GCG program package (Devereux et al., 1984), BLASTP,BLASTN and FASTA (Altschul et al, 1990).

[0033] The expression functional fragment of a nucleic acid” as usedherein means the minimal nucleic acid which is necessary to encode afunctional protein (or polypeptide). For instance, in situations where anucleic acid is provided comprising at the 5′ end and at the 3′ end morenucleotides than the actual open reading frame, the invention alsorelates to fragments of the nucleic acid which are smaller but whichstill contain the workable open reading frame. Also meant are parts ofthe open reading frame encoding a polypeptide having the same propertiesas the polypeptide encoded by the complete open reading frame.

[0034] The expression “a pathway eventually leading to programmed celldeath” refers to a sequence of steps ultimately leading to cell deathand which can be triggered at various steps in this pathway by variousagents, such as Bax, Bak, CED4, hydrogen peroxide, diamide and farnesol.The nucleic acid sequences to be used according to this aspect of theinvention from Saccharomyces cerevisiae are defined in SEQ ID NOs 17,19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53,55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89,91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119,121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147,149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175,177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203,205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231,233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259,261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287,289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315,317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343,345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371,373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 691, 693,695, 697, 699, 701, 703, 705, 707, 709, 711, 713 and 715; from Candidaalbicans are defined in SEQ ID NOs 397, 399, 401, 403, 405, 407, 409,411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437,439, 441, 443, 445, 447, 449, 451, 453, 455, 457, 459, 461, 463, 465,467, 469, 471, 473, 475, 477, 479, 481, 483, 485, 487, 489, 491, 493,495, 497, 499, 501, 503, 505, 507, 509, 511, 513, 515, 517, 519, 521,523, 525, 527, 529, 531, 533, 535, 537, 539, 541, 543, 545, 547, 549,551, 553, 555, 557, 559, 561, 563, 565, 567, 569, 571, 573, 575, 577,579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605,607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633,635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661,663, 665, 667, 669, 671, 673, 687, 718, 720, 722, 724, 726, 728, 730 and732.

[0035] The yeast or fungi according to the invention may be, but are notrestricted to, pathogenic yeast or fungi. As such, yeast or fungi maycause infections in healthy individuals as well as in immunocompromisedpatients.

[0036] The expression “treating diseases associated with yeast andfungi” not only refers to diseases or infections caused by saidorganisms but also refers to allergic reactions caused by saidorganisms, such as the so-called “professional diseases” in, forinstance, bakery and brewery and that are caused by yeast or fungi whichare commonly known as “non-pathogenic”. Some examples of specificdiseases associated with yeast or fungi are further exemplified.

[0037] The invention further relates to the use of nucleic acid sequencehomologues of SEQ ID NOs 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39,41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75,77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109,111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137,139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165,167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193,195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221,223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249,251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277,279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305,307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333,335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361,363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389,391, 393, 395, 397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417,419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445,447, 449, 451, 453, 455, 457, 459, 461, 463, 465, 467, 469, 471, 473,475, 477, 479, 481, 483, 485, 487, 489, 491, 493, 495, 497, 499, 501,503, 505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525, 527, 529,531, 533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557,559, 561, 563, 565, 567, 569, 571, 573, 575, 577, 579, 581, 583, 585,587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613,615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635, 637, 639, 641,643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669,671, 673, 687, 691, 693, 695, 697, 699, 701, 703, 705, 707, 709, 711,713, 715, 717, 719, 721, 723, 725, 727, 729 and 731 but isolated fromother yeast and fungi strains which are also involved in a pathwayeventually leading to programmed cell death. According to a morespecific embodiment, these nucleic acid sequences are derived fromAspergillus fumigatus.

[0038] In a more specific embodiment the invention relates to a nucleicacid encoding a polypeptide which is involved in a pathway eventuallyleading to programmed cell death of yeast or fungi selected from:

[0039] (a) a nucleic acid encoding a protein having an amino acidsequence as represented in any of SEQ ID NOs 398, 400, 402, 404, 406,408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434,436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462,464, 466, 468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490,492, 494, 496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518,520, 522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546,548, 550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 560, 562, 564,566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592,594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620,622, 624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648,650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 688,718, 720, 722, 724, 726, 728, 730 and 732, or encoding a functionalequivalent, derivative or bioprecursor of said protein;

[0040] (b) a nucleic acid encoding a protein having an amino acidsequence which is more than 70% similar, preferably more than 75% or 80%similar, more preferably more than 85%, 90% or 95% similar and mostpreferably more than 97% similar to any of the amino acid sequences asrepresented by any of SEQ ID NOs 398, 400, 402, 404, 406, 408, 410, 412,414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440,442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468,470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496,498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524,526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552,554, 556, 558, 560, 562, 564, 566, 568, 560, 562, 564, 566, 568, 570,572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598,600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626,628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654,656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 688, 718, 720, 722,724, 726, 728, 730 and 732,

[0041] (c) a nucleic acid encoding a protein having an amino acidsequence which is more than 70% identical, preferably more than 75% or80% identical, more preferably more than 85%, 90% or 95% identical andmost preferably more than 97% identical to any of the amino acidsequences as represented by any of SEQ ID NOs 398, 400, 402, 404, 406,408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434,436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462,464, 466, 468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490,492, 494, 496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518,520, 522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546,548, 550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 560, 562, 564,566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592,594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620,622, 624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648,650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 688,718, 720, 722, 724, 726, 728, 730 and 732,

[0042] (d) a nucleic acid comprising a sequence as represented in any ofSEQ ID 397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421,423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, 449,451, 453, 455, 457, 459, 461, 463, 465, 467, 469, 471, 473, 475, 477,479, 481, 483, 485, 487, 489, 491, 493, 495, 497, 499, 501, 503, 505,507, 509, 511, 513, 515, 517, 519, 521, 523, 525, 527, 529, 531, 533,535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557, 559, 561,563, 565, 567, 569, 571, 573, 575, 577, 579, 581, 583, 585, 587, 589,591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617,619, 621, 623, 625, 627, 629, 631, 633, 635, 637, 639, 641, 643, 645,647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673,687, 717, 719, 721, 723, 725, 727, 729 and 731;

[0043] (e) a nucleic acid which is more than 70% identical, preferablymore than 75% or 80% identical, more preferably more than 85%, 90% or95% identical and most preferably more than 97% identical to any of thenucleic acid sequences as represented by any of SEQ ID NO 397, 399, 401,403, 405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429,431, 433, 435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457,459, 461, 463, 465, 467, 469, 471, 473, 475, 477, 479, 481, 483, 485,487, 489, 491, 493, 495, 497, 499, 501, 503, 505, 507, 509, 511, 513,515, 517, 519, 521, 523, 525, 527, 529, 531, 533, 535, 537, 539, 541,543, 545, 547, 549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569,571, 573, 575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597,599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625,627, 629, 631, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653,655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 687, 717, 719, 721,723, 725, 727, 729 and 731,

[0044] (f) a nucleic acid encoding a functional fragment of any of thenucleic acid sequences as specified in a) to e), and,

[0045] (g) the complement of any of the nucleic acids as specified in a)to f).

[0046] In a preferred embodiment the invention relates to nucleic acidsfrom Candida albicans, as represented by the SEQ ID NOs 397, 399, 401,403, 405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429,431, 433, 435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457,459, 461, 463, 465, 467, 469, 471, 473, 475, 477, 479, 481, 483, 485,487, 489, 491, 493, 495, 497, 499, 501, 503, 505, 507, 509, 511, 513,515, 517, 519, 521, 523, 525, 527, 529, 531, 533, 535, 537, 539, 541,543, 545, 547, 549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569,571, 573, 575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597,599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625,627, 629, 631, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653,655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 687, 717, 719, 721,723, 725, 727, 729 and 731.

[0047] In an even more preferred embodiment the invention relates to anisolated nucleic acid from mammal or human origin which nucleic acidcorresponds to a mammal or human homologue of at least one of thesequences represented in SEQ ID NOs 17, 19, 21, 23, 25, 27, 29, 31, 33,35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69,71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103,105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131,133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159,161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187,189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215,217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243,245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271,273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299,301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327,329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355,357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383,385, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407, 409, 411,413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439,441, 443, 445, 447, 449, 451, 453, 455, 457, 459, 461, 463, 465, 467,469, 471, 473, 475, 477, 479, 481, 483, 485, 487, 489, 491, 493, 495,497, 499, 501, 503, 505, 507, 509, 511, 513, 515, 517, 519, 521, 523,525, 527, 529, 531, 533, 535, 537, 539, 541, 543, 545, 547, 549, 551,553, 555, 557, 559, 561, 563, 565, 567, 569, 571, 573, 575, 577, 579,581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607,609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635,637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663,665, 667, 669, 671, 673, 687, 691, 693, 695, 697, 699, 701, 703, 705,707, 709, 711, 713, 715, 717, 719, 721, 723, 725, 727, 729 and 731.

[0048] Therefore, according to a further preferred embodiment, theinvention relates to an isolated nucleic acid from mammal or humanorigin which nucleic acid sequence is selected from:

[0049] (a) a nucleic acid encoding a protein having an amino acidsequence as represented in any of SEQ ID NOs 676, 678, 680, 682, 684 and686, or encoding a functional equivalent, derivative or bioprecursor ofsaid protein;

[0050] (b) a nucleic acid encoding a protein having an amino acidsequence which is more than 70% similar, preferably more than 75% or 80%similar, more preferably more than 85%, 90% or 95% similar and mostpreferably more than 97% similar to any of the amino acid sequences asrepresented by any of SEQ ID NOs 676, 678, 680, 682, 684 and 686;

[0051] (c) a nucleic acid encoding a protein having an amino acidsequence which is more than 70% identical, preferably more than 75% or80% identical, more preferably more than 85%, 90% or 95% identical andmost preferably more than 97% identical to any of the amino acidsequences as represented by any of SEQ ID NOs 676, 678, 680, 682, 684and 686;

[0052] (d) a nucleic acid comprising a sequence as represented in any ofSEQ ID NOs 675, 677, 679, 681, 683 and 685;

[0053] (e) a nucleic acid which is more than 70% identical, preferablymore than 75 or 80% identical, more preferably more than 85%, or 90% or95% identical and most preferably more than 97% identical to any of thenucleic acid sequences as represented by any of SEQ ID NOs 675, 677,679, 681, 683 and 685;

[0054] (f) a nucleic acid encoding a functional fragment of any of thenucleic acids as specified in a) to e); and

[0055] (g) the complement of any of the nucleic acids as specified in a)to f),

[0056] for the preparation of a medicament for treating diseasesassociated with yeast or fungi.

[0057] The invention also relates to the use of said nucleic acids fortreating and/or preventing and/or alleviating proliferative disorders orfor the prevention of apoptosis in certain disorders or diseases.

[0058] The expression “proliferative disorders” or “proliferativediseases” refers to an abnormality within a patient or animal such ascancer. Normal cells start to proliferate due to a change in the codingor non-coding sequence of the DNA resulting in a swollen or distendedtissue. Mutation may arise without obvious cause. An abnormal benign ormalignant mass of tissue is formed that is not inflammatory. Cells ofpre-existent tissue start to divide unexpectedly and resulting cell masspossesses no physiologic function.

[0059] The expression “apoptosis” or “apoptosis-related diseases”includes diseases such as autoimmunity diseases, ischemia, diseasesrelated with viral infections or neurodegenerations.

[0060] It should be clear that the invention also relates to all nucleicacids according to the invention and which are specifically describedabove, and which can be DNA, cDNA, genomic DNA, synthetic DNA, or RNAwherein T is replaced by U. A nucleic acid according to the inventionmay also comprise any modified nucleotide known in the art.

[0061] The term “nucleic acid sequence” also includes the complementarysequence to any single stranded sequence given.

[0062] According to the invention, these sequences and their homologuesin other yeast and fungi or in human or other mammals as well as thepolypeptides which they encode represent novel molecular targets whichcan be incorporated into an assay to selectively identify compoundscapable of inhibiting or activating expression of such polypeptides.Furthermore, the invention also relates to the potential use of saidsequences in alleviating diseases or conditions associated with yeast orfungi infections, such as diseases caused by Candida spp., Aspergillusspp., Microsporum spp., Trichophyton spp., Fusarium spp., Zygomycetesspp., Botritis spp., Cladosporium spp., Malassezia spp., Epidermophytonfloccosum, Blastomyces dermatitidis, Coccidioides immitis, Histoplasmacapsulatum, Paracoccidioides brasiliensis, Cryptococcus neoformans, andSporothrix schenckii, such as, but not limited to:

[0063] Candidiasis, caused by C. albicans and other members of the genusCandida, which are primary or secondary mycotic infections, also namedcandidosis, moniliasis and thrush;

[0064] Aspergilliosis, caused by members of the genus Aspergillus, forma spectrum of diseases;

[0065] Histoplasmosis, caused by Histoplasma capsulatum, which is apulmonary disease always seen in HIV positive or other immunocompromisedindividuals;

[0066] Paracoccidioidomycosis, caused by Paracoccidioides brasiliensis,which is a granulomatous disease that originates as a pulmonary disease;

[0067] Blastomycosis, caused by Blastomyces dermatitidis, which may be abenign and self-limiting infection or a chronic granulomatous andsuppurative mycosis, also named Chicago disease or Gilchrist's disease;

[0068] Coccidioidomycosis, caused by Coccidioides imminitis, and whichis a respiratory infection that typically resolves rapidly, but themycosis can become acute, chronic, severe or fatal; also named SanJoaquin Valley fever or Valley fever;

[0069] Cryptococcosis, caused by Cryptococcus neoformans, which is achronic, subacute to acute pulmonary, systemic or meningitic disease,also named Torulosis;

[0070] Sporotrichosis, caused by Sporothrix schenckii, which is achronic infection characterized by nodular lesions of cutaneous orsubcutaneous tissues and adjacent lymphatics that suppurate, ulcerateand drain.

[0071] Some of the pathways leading to apoptosis are conserved betweenmammalian cells and yeast or fungi. Therefore the invention also relatesto the potential use of homologous sequences from human or mammalianorigin for preventing and/or alleviating diseases or conditions whereapoptosis or non-apoptosis of cells is impaired, for instance inproliferative disorders. In this respect also cancer can be seen as aproliferative disorder. Furthermore, targets which are part of such aconserved pathway may be used to stimulate or inhibit the apoptosis inmammalian cells. E.g. stimulation of apoptosis is desirable in thetreatment of tumor cells/tissues.

[0072] Human homologues according to the invention can be obtained byselective hybridisation of the yeast and candida nucleic acid moleculesof the invention against human genome or cDNA libraries according tomethods well known in the art (Sambrook et al., 1989). Human polypeptidehomologues are obtained from the corresponding human nucleic acidhomologous nucleotide sequences.

[0073] The present invention further relates to a nucleic acid capableof selectively hybridising to at least one of the nucleic acid moleculesaccording to the invention, or the complement thereof.

[0074] The term “selectively hybridising” or “specifically hybridising”means hybridising under conditions wherein sequences can be detectedwhich are homologues of the sequences of the invention, but which arefor instance derived from heterologous cells or organisms, and whereinsaid sequences do not hybridize with known sequences. In a preferredembodiment, mammalian homologues can be detected. It is well known tothe person skilled in the art which methods for hybridisation can beused and which conditions are necessary for selectively or specificallyhybridising. Preferably, hybridization under high stringency conditionscan be applied (Sambrook et al., 1989).

[0075] As such, the present invention also relates to the use of thenucleic acid sequences of the invention for detecting homologues inheterologous organisms including but not limited to mammalian organisms.

[0076] The invention also relates to an isolated nucleic acid comprisinga human homologue of at least one of the yeast or candida nucleic acidsdescribed earlier. The invention also relates to a polypeptide encodableby said human homologue of said nucleic acid.

[0077] In a further embodiment the invention also relates to anexpression vector comprising a human homologue of at least one of theyeast or candida nucleic acids described herein. Said expression vectoraccording can be an expression vector wherein said nucleic acid sequenceis operably linked to one or more control sequences allowing theexpression in prokaryotic and/or eukaryotic host cells. According to afurther embodiment, the expression vector comprises an induciblepromoter and/or a reporter molecule.

[0078] The invention also relates to a host cell transformed,transfected or infected with any of the above described vectors.

[0079] According to a preferred embodiment, the invention relates to anantisense version of any of the nucleic acids of the invention anddescribed above.

[0080] The present invention more particularly relates to an antisensemolecule comprising a nucleic acid capable of selectively hybridising toat least one of the nucleic acids of the invention. In an interestingembodiment the invention relates to a nucleic acid capable ofselectively hybridising to a human homologue of at least one yeast orcandida nucleic acid described herein.

[0081] Polynucleotides according to the invention may be inserted intovectors in an antisense orientation in order to provide for theproduction of antisense RNA. Antisense RNA or other antisense nucleicacids may also be produced by synthetic means.

[0082] The present invention also advantageously provides nucleic acidmolecules of at least approximately 10 contiguous nucleotides of anucleic acid according to the invention and preferably from 10 to 50nucleotides. These sequences may, advantageously be used as probes orprimers to initiate replication, or the like. Such nucleic acidsequences may be produced according to techniques well known in the art,such as by recombinant or synthetic means. The probes will hybridisespecifically with any of the nucleic acid molecules of the invention.The primers will specifically amplify any of the nucleic acid moleculesof the invention. The probes or primers according to the invention mayalso be used in diagnostic kits or the like for detecting the presenceof a nucleic acid according to the invention. These tests generallycomprise contacting the probe with the sample under hybridisingconditions and detecting the presence of any duplex or triplex formationbetween the probe and any nucleic acid in the sample.

[0083] According to the present invention these probes may be anchoredto a solid support. Preferably, they are present on an array so thatmultiple probes can simultaneously hybridize to a single biologicalsample. The probes can be spotted onto the array or synthesized in situon the array. (Lockhart et al., 1996). A single array can contain morethan 100, 500 or even 1,000 different probes in discrete locations. Sucharrays can be used to screen for compounds interacting with said probes.

[0084] Advantageously, the nucleic acid sequences, according to theinvention may be produced using recombinant or synthetic means, such asfor example using PCR cloning mechanisms which generally involve makinga pair of primers, which may be from approximately 10 to 50 nucleotidesto a region of the gene which is desired to be cloned, bringing theprimers into contact with mRNA, cDNA, or genomic DNA from the yeast orfungal cell, performing a polymerase chain reaction under conditionswhich bring about amplification of the desired region, isolating theamplified region or fragment and recovering the amplified DNA.Generally, such techniques as defined herein are well known in the art,such as described in Sambrook et al. (1989). These techniques can beused to clone homologues of the nucleic acid sequences of the inventionin other organisms.

[0085] The nucleic acids or oligonucleotides according to the inventionmay carry a revealing label. Suitable labels include radioisotopes suchas ³²P, ³³P or ³⁵S, enzyme labels or other protein labels such as biotinor fluorescent markers. Such labels may be added to the nucleic acids oroligonucleotides of the invention and may be detected using techniquesknown in the art.

[0086] According to another embodiment of the invention, the nucleicacid sequences according to the invention as defined above may,advantageously, be included in a suitable vector, preferably anexpression vector which may be transformed, transfected or infected intoa host cell. In such an expression vector the nucleic acid is operablylinked to one or more control sequences allowing the expresssison inhost cells, such as a suitable promotor, or the like, to ensureexpression of the proteins according to the invention in a suitableprokaryotic or eukaryotic host cell. Said promoter may be eitherconstitutive, inducible or cell- or tissue- or organ-specific. Theexpression vector may advantageously be a plasmid, cosmid, virus orother suitable vector which is known to those skilled in the art. Theexpression vector and the host cell defined herein also form part of thepresent invention. Said host cell can be from bacterial, yeast, fungal,insect, mammal or human origin, or any other host wherein said vectorcan be introduced by at least one of the methods known in the art.However, preferred host cells are lower eukaryotic cells such as a yeastcell or a fungal cell. Yeast and fungal cells are particularlyadvantageous because they provide the necessary post-translationalmodifications to the expressed proteins of the invention, similar tothose of the natural proteins from which they are derived. Thesemodifications confer optimal conformation of said proteins, which whenisolated may advantageously be used in kits, methods or the like.

[0087] In a further embodiment, the expression vector may furthercomprise an inducible promoter, and/or further a reporter molecule.

[0088] The invention further relates to any one of the nucleic acids asdefined above for use as a medicament.

[0089] Nucleotide sequences according to the invention are particularlyadvantageous for providing selective therapeutic targets for treatingyeast or fungi-associated infections. For example, an antisense nucleicacid capable of binding to the nucleic acid sequences according to theinvention may be used to selectively inhibit expression of thecorresponding polypeptides, leading to impaired growth or death of yeastand fungi with reductions of associated illnesses or diseases.

[0090] Also envisaged in the present invention are promoter or othercontrol sequences that are comprised within the nucleic acids of theinvention, said nucleic acid control sequences can also serve as atarget for the identification of compounds or proteins which interferewith the control of expression of downstream encoded polypeptides.

[0091] Furthermore, also the human homologues of the yeast and candidanucleic acids may be useful in diseases where apoptosis of cells plays asubstantial role, both in situations where apoptosis of (particular)cells is wanted or unwanted.

[0092] The invention thus also relates to the use of any of the nucleicacids of the invention or to a human homologue thereof for treatingproliferative disorders or for the prevention of apoptosis in certaindisorders or diseases. As described above, the invention also relates tothe use of antisense molecules of the nucleic acids of the invention orto an antisense of any of the human homologues for treatingproliferative disorders or for the prevention of apoptosis in certaindisorders or diseases.

[0093] Said nucleic acids, human homologues and antisense molecules canalso be used for the preparation of a medicament for treating orpreventing the above-mentioned diseases.

[0094] According to yet another embodiment, the invention relates to atleast one polypeptide encodable by a nucleic acid of the invention.

[0095] The invention also relates to the use of a polypeptide which isinvolved in a pathway eventually leading to programmed cell death ofyeast or fungi, said polypeptide being selected from:

[0096] (a) a protein having an amino acid sequence as represented in anyof SEQ ID NOs 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44,46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80,82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112,114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140,142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168,170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196,198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224,226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252,254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280,282, 284, 286, 288, 290, 292, 294, 296, 298, 290, 292, 294, 296, 298,300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326,328, 330, 332, 324, 326, 328, 340, 342, 344, 346, 348, 350, 352, 354,356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382,384, 386, 388, 390, 392, 394, 396, 398, 400, 402, 404, 406, 408, 410,412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438,440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466,468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494,496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522,524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550,552, 554, 556, 558, 560, 562, 564, 566, 568, 560, 562, 564, 566, 568,570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596,598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624,626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652,654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 688, 692, 694,696, 698, 700, 702, 704, 706, 708, 710, 712, 714, 716, 718, 720, 722,724, 726, 728, 730 and 732, or encoding a functional equivalent,derivative or bioprecursor of said protein;

[0097] (b) a protein having an amino acid sequence which is more than70% similar, preferably more than 75% or 80% similar, more preferablymore than 85%, 90% or 95% similar and most preferably more than 97%similar to any of the amino acid sequences as represented by any of SEQID NOs 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48,50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84,86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116,118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144,146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172,174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200,202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228,230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256,258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284,286, 288, 290, 292, 294, 296, 298, 290, 292, 294, 296, 298, 300, 302,304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330,332, 324, 326, 328, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358,360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, 384, 386,388, 390, 392, 394, 396, 398, 400, 402, 404, 406, 408, 410, 412, 414,416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442,444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468, 470,472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496, 498,500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526,528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554,556, 558, 560, 562, 564, 566, 568, 560, 562, 564, 566, 568, 570, 572,574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600,602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628,630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656,658, 660, 662, 664, 666, 668, 670, 672, 674, 688, 692, 694, 696, 698,700, 702, 704, 706, 708, 710, 712, 714, 716, 718, 720, 722, 724, 726,728, 730 and 732,

[0098] (c) a protein having an amino acid sequence which is more than70% identical, preferably more than 75% or 80% identical, morepreferably more than 85%, 90% or 95% identical and most preferably morethan 97% identical to any of the amino acid sequences as represented byany of SEQ ID NOs 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42,44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78,80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110,112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138,140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166,168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194,196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222,224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250,252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278,280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 290, 292, 294, 296,298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324,326, 328, 330, 332, 324, 326, 328, 340, 342, 344, 346, 348, 350, 352,354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380,382, 384, 386, 388, 390, 392, 394, 396, 398, 400, 402, 404, 406, 408,410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436,438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464,466, 468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492,494, 496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520,522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548,550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 560, 562, 564, 566,568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594,596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622,624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650,652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 688, 692,694, 696, 698, 700, 702, 704, 706, 708, 710, 712, 714, 716, 718, 720,722, 724, 726, 728, 730 and 732, and,

[0099] (d) a functional fragment of any of said proteins as defined ina) to c),

[0100] for the preparation of a medicament for treating diseasesassociated with yeast or fungi.

[0101] The term “functional fragment” of a protein means a truncatedversion of the original protein or polypeptide referred to. Thetruncated protein sequence can vary widely in length; the minimum sizebeing a sequence of sufficient size to provide a sequence with at leasta comparable function and/or activity of the original sequence referredto, while the maximum size is not critical. In some applications, themaximum size usually is not substantially greater than that required toprovide the desired activity and/or function(s) of the originalsequence. A functional fragment can also relate to a subunit withsimilar function as said protein. Typically, the truncated amino acidsequence will range from about 5 to about 60 amino acids in length. Moretypically, however, the sequence will be a maximum of about 50 aminoacids in length, preferably a maximum of about 60 amino acids. It isusually desirable to select sequences of at least about 10, 12 or 15amino acids.

[0102] Functional fragments include those comprising an epitope which isspecific or unique for the proteins according to the invention. Epitopesmay be determined using, for example, peptide scanning techniques asdescribed in Geysen et al. (1986). Preferred functional fragments have alength of at least, for example, 5, 10, 25, 50, 75, 100, 125, 150, 175or 200 amino acids.

[0103] The polypeptides to be used according to the invention fromSaccharomyces cerevisiae, are represented by SEQ ID NOs 18, 20, 22, 24,26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60,62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96,98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124,126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152,154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180,182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208,210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236,238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264,266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292,294, 296, 298, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310,312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 324, 326, 328,340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366,368, 370, 372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394,396, 692, 694, 696, 698, 700, 702, 704, 706, 708, 710, 712, 714 and 716.Also according to the invention is the use of the polypeptides fromCandida albicans as represented by the SEQ ID NOs 398, 400, 402, 404,406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432,434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460,462, 464, 466, 468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488,490, 492, 494, 496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516,518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544,546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 560, 562,564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590,592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618,620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646,648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674,688, 718, 720, 722, 724, 726, 728, 730 and 732, and the use of humanpolypeptides as represented by SEQ ID NOs 676, 678, 680, 682, 684 and686.

[0104] Thus, according to a preferred embodiment, the present inventionrelates to an isolated polypeptide which is involved in a pathway forprogrammed cell death of yeast or fungi, for instance a Candida spp.,selected from:

[0105] (a) a polypeptide having an amino acid sequence as represented inany of SEQ ID NOs 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418,420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446,448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474,476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502,504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530,532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558,560, 562, 564, 566, 568, 560, 562, 564, 566, 568, 570, 572, 574, 576,578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604,606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632,634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660,662, 664, 666, 668, 670, 672, 674, 688, 718, 720, 722, 724, 726, 728,730 and 732, or encoding a functional equivalent, derivative orbioprecursor of said protein;

[0106] (b) a polypeptide having an amino acid sequence which is morethan 70% similar, preferably more than 75% or 80% similar, morepreferably more than 85%, 90% or 95% similar and most preferably morethan 97% similar to any of the amino acid sequences as represented byany of SEQ ID NOs 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418,420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446,448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474,476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502,504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530,532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558,560, 562, 564, 566, 568, 560, 562, 564, 566, 568, 570, 572, 574, 576,578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604,606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632,634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660,662, 664, 666, 668, 670, 672, 674, 688, 718, 720, 722, 724, 726, 728,730 and 732,

[0107] (c) a polypeptide having an amino acid sequence which is morethan 70% identical, preferably more than 75% or 80% identical, morepreferably more than 85%, 90% or 95% identical and most preferably morethan 97% identical to any of the amino acid sequences as represented byany of SEQ ID NOs 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418,420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446,448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474,476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502,504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530,532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558,560, 562, 564, 566, 568, 560, 562, 564, 566, 568, 570, 572, 574, 576,578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604,606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632,634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660,662, 664, 666, 668, 670, 672, 674, 688, 718, 720, 722, 724, 726, 728,730 and 732, and

[0108] (d) a functional fragment of any of said polypeptides as definedin a) to c).

[0109] According to a further preferred embodiment, the presentinvention relates to an isolated polypeptide which is involved in apathway for programmed cell death of mammalian cells selected from:

[0110] (a) a polypeptide having an amino acid sequence as represented inany of SEQ ID NOs 676, 678, 680, 682, 684 and 686, or encoding afunctional equivalent, derivative or bioprecursor of said protein;

[0111] (b) a polypeptide having an amino acid sequence which is morethan 70% similar, preferably more than 75% or 80% similar, morepreferably more than 85%, 90% or 95% similar and most preferably morethan 97% similar to any of the amino acid sequences as represented byany of SEQ ID NOs human 676, 678, 680, 682, 684 and 686;

[0112] (c) a polypeptide having an amino acid sequence which is morethan 70% identical, preferably more than 75% or 80% identical, morepreferably more than 85%, 90% or 95% identical and most preferably morethan 97% identical to any of the amino acid sequences as represented byany of SEQ ID NOs 676, 678, 680, 682, 684 and 686; and,

[0113] (d) a functional fragment of any of said polypeptides as definedin a) to c).

[0114] The invention also relates to the polypeptides of the inventionand described above for use as a medicament.

[0115] Pharmaceutical or fungicidal compositions comprising at least oneof the nucleic acids, antisense molecules, polypeptides of the inventionoptionally together with a pharmaceutically acceptable carrier, diluentor excipient therefor, are also part of the invention.

[0116] The polypeptides described above or the human or mammalhomologues thereof can also be used for treating proliferative disordersor for the prevention of apoptosis in certain diseases.

[0117] The invention furthermore relates to a pharmaceutical compositionfor use as a medicament for treating proliferative disorders or for theprevention of apoptosis in certain diseases comprising a nucleic acidmolecule of the invention or a human homologue thereof, an antisensemolecule to at least one of the nucleic acids of the invention or anantisense molecule to a mammalian homologue of said nucleic acid or apolypeptide of the invention or a human homologue thereof together witha pharmaceutically acceptable carrier, diluent or excipient therefor.

[0118] The polypeptide or protein according to the invention may alsoinclude variants of any of the polypeptides of the invention asspecified above having conservative amino acid changes.

[0119] The present invention also relates to a vaccine for immunizing amammal comprising at least one (recombinant) nucleic acid molecule or atleast one (recombinant) polypeptide of the invention in apharmaceutically acceptable carrier. Preferred vaccines are those thatcan be used for immunization against infections caused by yeast andfungi. Other preferred vaccines can be used for immunizing mammalsagainst proliferative disorders or for preventing apoptosis in certaindiseases.

[0120] Pharmaceutically acceptable carriers include any carrier thatdoes not itself induce the production of antibodies harmful to theindividual receiving the composition. Suitable carriers are typicallylarge, slowly metabolizing macromolecules such as proteins,polysaccharides, polylactic acids, polyglycolic acids, polymeric aminoacids, amino acid copolymers; and inactive virus particles. Suchcarriers are well known to those of ordinary skill in the art.

[0121] A “vaccine” is an immunogenic composition capable of elicitingprotection against infections caused by yeast or fungi, whether partialor complete.

[0122] Said vaccine compositions may include prophylactic as well astherapeutic vaccine compositions. When a vaccine is used for protectingindividuals against certain infections or diseases, it is called aprophylactic vaccine. A vaccine may also be useful for treatment of anindividual, in which case it is called a therapeutic vaccine.

[0123] The term “therapeutic” refers to a composition capable oftreating infections caused by yeast or fungi or capable of treatingproliferative disorders.

[0124] Also encompassed within the present invention are antibodies,monoclonal or polyclonal, capable of specifically binding to one or moreepitopes of the polypeptides or proteins of the invention. Thepolypeptides of the invention are represented in SEQ ID NOs 18, 20, 22,24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58,60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94,96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124,126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152,154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180,182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208,210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236,238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264,266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292,294, 296, 298, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310,312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 324, 326, 328,340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366,368, 370, 372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394,396, 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422,424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450,452, 454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478,480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506,508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534,536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562,564, 566, 568, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580,582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608,610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636,638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664,666, 668, 670, 672, 674, 676, 678, 680, 682, 684, 686, 688, 692, 694,696, 698, 700, 702, 704, 706, 708, 710, 712, 714, 716, 718, 720, 722,724, 726, 728, 730 and 732.

[0125] The term “specific binding” implies that there is substantiallyno cross-reaction of the antibody with other proteins.

[0126] The antibodies according to the invention may be producedaccording to techniques which are known to those skilled in the art.Monoclonal antibodies may be prepared using conventional hybridomatechnology as described by Kohler and Milstein (1979). Polyclonalantibodies may also be prepared using conventional technology well knownto those skilled in the art, and which comprises inoculating a hostanimal, such as a mouse, with a protein or epitope according to theinvention and recovering the immune serum. The present invention alsoincludes fragments of whole antibodies which maintain their bindingactivity, such as for example, Fv, F(ab′) and F(ab′)₂ fragments as wellas single chain antibodies.

[0127] The antibodies of the invention are capable of specificallybinding to at least one of the yeast or candida polypeptides as definedearlier or to a human homologue thereof or to a specific epitope of saidpolypeptide or said human homologue. The invention also relates to theuse of said antibodies in treating and/or preventing and/or alleviatingproliferative disorders or for the prevention of apoptosis in certaindiseases. Said antibodies may also be used for the preparation of amedicament for and/or preventing and/or alleviating proliferativedisorders or for the prevention of apoptosis in certain diseases.

[0128] Antibodies according to the invention may also be used in amethod of detecting the presence of a polypeptide according to theinvention, which method comprises reacting the antibody with a sampleand identifying any protein bound to said antibody. A kit may also beprovided for performing said method which comprises an antibodyaccording to the invention and means for reacting the antibody with saidsample.

[0129] The antibodies according to the invention may be used as amedicament or may be comprised in a pharmaceutical composition.According to a more specific embodiment, the antibodies may be used inthe preparation of a medicament for treating diseases associated withyeast and fungi where the yeast or fungus is chosen from, but notrestricted to Candida spp., Aspergillus spp., Microsporum spp.,Trichophyton spp., Fusarium spp., Zygomycetes spp., Botritis, spp.,Cladosporium spp., Malassezia spp., Epidermophyton floccosum,Blastomyces dermatitidis, Coccidioides imminitis, Histoplasmacapsulatum, Paracoccidioides brasiliensis, Cryptococcus neoformans, andSporothrix schenckii.

[0130] The invention also relates to a method of preventing infectionwith yeast or fungi, comprising administering a composition containingat least one polypeptide of the invention to a mammal in effectiveamount to stimulate the production of protective antibody or protectiveT-cell response.

[0131] According to another embodiment, the present invention provides amethod of identifying compounds or polypeptides which selectivelyinhibit, induce or interfere with the expression/production of thepolypeptides encoded by the nucleotide sequences of the Invention, orcompounds which selectively inhibit, activate or interfere with thefunctionality of polypeptides expressed from the nucleotide sequencesaccording to the invention, or which selectively inhibit, induce orinterfere with the metabolic pathways in which these polypeptides areinvolved. Compounds (or polypeptides) may carry agonistic orantagonistic properties. The compounds (and polypeptides) to be screenedmay be of extracellular, intracellular, biologic or chemical origin.

[0132] Different alternative methods for identification of saidcompounds or polypeptides form part of the present invention.

[0133] According to a specific embodiment the invention relates to amethod of identifying compounds which selectively modulate expression orfunctionality of polypeptides involved in a pathway eventually leadingto programmed cell death of yeast and fungi or in metabolic pathways inwhich said polypeptides are involved, which method comprises (a)contacting a compound to be tested with yeast or fungal cellstransformed, transfected or infected with an expression vectorcomprising an antisense sequence of at least one of the nucleic acidsequences of the invention, which expression results in underexpressionof said polypeptide, in addition to contacting one or more wild typecells with said compound, (b) monitoring the growth and/or death rate oractivity of said transformed, transfected or infected cells compared tosaid wild type cells; wherein differential growth or activity of saidtransformed, transfected or infected yeast or fungal cells is indicativeof selective action of said compound on a polypeptide in the same or aparallel pathway, (c) alternatively monitoring the growth and/or deathrate and/or activity of said transformed, transfected or infected cellscompared to transformed, transfected or infected cells which were notcontacted with the compound to be tested, wherein differential growth oractivity of said mutated yeast or fungi cells is indicative of selectiveaction of said compound on a polypeptide in the same or a parallelpathway, (d) alternatively monitoring changes in morphologic and/orfunctional properties of components in said transformed, transfected orinfected cells caused by the addition of the compound to be tested, and(e) optionally identifying the compound.

[0134] Alternative methods for identifying compounds which selectivelymodulate expression or functionality of polypeptides involved in apathway eventually leading to programmed cell death of yeast or fungi orin metabolic pathways in which said compounds are involved, may comprisethe use of any other method known in the art resulting in geneactivation, gene inactivation, gene modulation or gene silencing.

[0135] Another alternative to the above described method comprises (a)contacting a compound to be tested with a genetically modified yeast orfungus in which modification results in the overexpression orunderexpression of at least one of the nucleic acids or the polypeptidesof the invention, which overexpression or underexpression of saidnucleic acid or polypeptide prevents, delays or sensitizes for apoptosisof said genetically modified yeast or fungus, in addition to contactingwild type cells with said compound, (b) monitoring the growth and/ordeath rate and/or activity of said genetically modified yeast or fungicells compared to said wild type cells wherein differential growth oractivity of said genetically modified yeast or fungi cells is indicativeof selective action of said compound on a polypeptide in the same or aparallel pathway, (c) alternatively monitoring the growth and/or deathrate and/or activity of said genetically modified cells compared togenetically modified cells which were not contacted with the compound tobe tested, wherein differential growth or activity of said geneticallymodified yeast of fungi cells is indicative of selective action of saidcompound on a polypeptide in the same or a parallel pathway, (d)alternatively monitoring changes in morphologic and/or functionalproperties of components in said genetically modified cells caused bythe addition of the compound to be tested, and, (e) optionallyidentifying the compound.

[0136] The invention also relates to a method of identifying compoundswhich selectively modulate expression of polypeptides which are involvedin a pathway eventually leading to programmed cell death of yeast orfungi which method comprises (a) contacting host cells transformed,transfected or infected with an expression vector comprising a promotersequence of a nucleic acid molecule of the invention joined in framewith a reporter gene and (b) monitoring increased or decreasedexpression of said reporter gene caused by the addition of the compoundbeing tested. This enables to analyse the influence of the compound ontoall/most aspects of transcriptional activation. Alternatively additionaltests can routinely be performed to test the influence of the compoundonto mRNA stability, translation and protein stability. All theseaspects influence the concentration of corresponding proteins andconsequently influence the effect of these on the metabolism of thecell.

[0137] The invention further relates to a method of identifyingcompounds or polypeptides which bind to or modulate the properties ofpolypeptides which are involved in a pathway eventually leading toprogrammed cell death of yeast or fungi, which method comprises (a)contacting a compound or polypeptide to be tested with at least one ofthe polypeptides of the invention, (b) detecting the complex formedbetween the compound or polypeptide to be tested and said polypeptide,(c) alternatively, examining the diminution/increase of complexformation between said polypeptide and a receptor/binding partner,caused by the addition of the compound or polypeptide being tested, (c)alternatively, examining the alteration in the functional activity ofthe polypeptide, caused by the addition of the compound or polypeptidebeing tested, and (d) optionally identifying the compound orpolypeptide.

[0138] The invention also relates to a method for identifying compoundsinteracting with a polypeptide involved in a pathway eventually leadingto programmed cell death of yeast and fungi comprising the steps of (a)providing a two-hybrid screening system wherein a polypeptide of theinvention and a protein interacting with said polypeptide or aninteracting polypeptide obtainable by a method as described above, areexpressed, (b) interacting said compound with the complex formed by theexpressed proteins as defined in a), (c) detecting a second complex,wherein the presence of said second complex identifies a compound whichspecifically binds to one of said polypeptide or to said second complex,and optionally (d) identifying the compound. According to anotherembodiment the invention relates to a method for identifying compoundswhich selectively modulate expression of polypeptides which are involvedin a pathway eventually leading to programmed cell death of yeast orfungi which method comprises: (a) contacting host cells transformed,transfected or infected with an expression vector comprising a promotersequence of a nucleic acid of the invention joined in frame with areporter gene, (b) monitoring increased or decreased expression of saidreporter gene caused by the addition of the compound being tested, and,optionally (c) identifying the compound.

[0139] Yet another embodiment of the invention is a method foridentifying polypeptides involved in a pathway eventually leading toprogrammed cell death comprising the steps of: (a) providing a twohybrid system wherein a polypeptide encoded by a nucleic acid or by anyof the vectors of the invention as a bait and a S. cerevisiae cDNAlibrary as a prey are used, (b) detecting an interaction between saidpolypeptide and a S. cerevisiae polypeptide encoded by said cDNAlibrary, and, optionally (c) identifying said S. cerevisiae polypeptide.

[0140] The term “cells” as used in the above methods relates to any typeof cells such as, but not limited to bacterial, yeast, fungal, plant orhuman cells.

[0141] Compounds found using this approach may additionally be tested ontheir efficiency in killing or inhibiting the growth of wild type cellsin order to confirm their utility as medicament for treating wild typepathogenic strains/tumor cells.

[0142] According to the invention, the term “mutation” includes pointmutations, deletions, insertions, duplications or any modification inthe nucleic acid encoding said polypeptide, or at a different locationin the genome of said cells, influencing the expression of said nucleicacid or polypeptide. In case point mutations occur, the number ofnucleotides will be identical compared to the original sequence; only achange in nucleotide sequence can be observed. This stands in contrastwith the other listed mutations where the number of the nucleotides willbe different from the number observed in the wild type sequence andconsequently will also reflect in a change of the nucleotide sequence.

[0143] Changes in morphologic and/or functional properties of cellcomponents which can be monitored include for example morphological andmolecular changes such as abnormal cell morphology, nuclearfragmentation, DNA breakage or changes in the expression of certainenzymes such as caspases, as well as monitoring changes in membranepotential or activity of mitochondria and release of cytochrome c frommitochondria. All these changes can be monitored on the whole cell whichis contacted to the compound to be tested.

[0144] Detection of the complex formation can be performed using severalapproaches. First, binding of a compound onto a polypeptide can bestudied using classical binding tests: one of the binding partners,compound or polypeptide is labeled and interaction of both is measured.Most of these tests comprise following steps: incubating both bindingpartners in conditions where binding is allowed, separation of freelabel from bound label present in the complex formed between bothpartners, and measuring the number of labeled complexes formed.Separation of free and bound label can be performed via filtration,centrifugation or other means as known by the person skilled in the art.Other techniques allow visualisation of complex formation without theneed of such a separating step. For example, test systems using SPA(scincillation proximity assay) beads are based on the principle thatradioactive ³H can only be measured when present in scincillation fluid.SPA beads contain scincillation fluid and can be coated with one of thebinding partners. When this bead is approached and binds the otherbinding partner which is radioactively labeled, a signal will bedetected allowing the complex to be visualised. Binding of theradioactive compound onto the scincillation bead is needed in order toresult in a detectable signal; non-bound radioactive partners that stayfree into the solution will not result in a detectable signal.

[0145] The protein or peptide fragments according to the inventionemployed in such a method may be for example in solution or coated onsuspended beads as described above. Alternatively, these can be affixedto a solid support, borne on a cell or phage surface or locatedintracellularly.

[0146] When protein or peptide fragments are coated on solid supports,they can be tested for their binding affinity for large numbers ofcompounds. These can be used in different kinds of high throughputscreenings in order to identify compounds having suitable bindingaffinity to the polypeptides according to the invention. Platformtechnologies or technologies based on SPR (see below) can be applied.

[0147] One may measure for example, the formation of complexes betweenthe proteins of the invention and the compound being tested.Alternatively, one may examine the diminution or increase of complexformation between the protein according to the invention and areceptor/binding partner caused by the compound being tested.

[0148] Proteins which interact with the polypeptide of the invention maybe identified by investigating protein-protein interactions using thetwo-hybrid vector system first proposed by Chien et al. (1991).

[0149] This technique is based on functional reconstitution in vivo of atranscription factor which activates a reporter gene. More particularlythe technique comprises providing an appropriate host cell with a DNAconstruct comprising a reporter gene under the control of a promoterregulated by a transcription factor having a DNA binding domain and anactivating domain, expressing in the host cell a first hybrid DNAsequence encoding a first fusion of a fragment or all of a nucleic acidsequence according to the invention and either said DNA binding domainor said activating domain of the transcription factor, expressing in thehost at least one second hybrid DNA sequence, such as a library or thelike, encoding putative binding proteins to be investigated togetherwith the DNA binding or activating domain of the transcription factorwhich is not incorporated in the first fusion; detecting any binding ofthe proteins to be investigated with a protein according to theinvention by detecting for the presence of any reporter gene product inthe host cell; optionally isolating second hybrid DNA sequences encodingthe binding protein.

[0150] An example of such a technique utilizes the GAL4 protein inyeast. Gal4 is a transcriptional activator of galactose metabolism inyeast and has a separate domain for binding to activators upstream ofthe galactose metabolising genes as well as a protein-binding domain.Nucleotide vectors may be constructed, one of which comprises thenucleotide residues encoding the DNA binding domain of Gal4. Thesebinding domain residues may be fused to a known protein encodingsequence, such as for example the nucleic acids according to theinvention. The other vector comprises the residues encoding theprotein-binding domain of Gal4. These residues are fused to residuesencoding a test protein. Any interaction between polypeptides encoded bythe nucleic acid according to the invention and the protein to be testedleads to transcriptional activation of a reporter molecule in a GAL4transcription deficient yeast cell into which the vectors have beentransformed. Preferably, a reporter molecule such as β-galactosidase isactivated upon restoration of transcription of the yeast galactosemetabolism genes. Alternatively, other reporter proteins can be usedsuch as EGFP (enhanced green fluorescent protein), or hEGFP. This latterhas a decreased lifetime enabling the system to screen for compoundsimproving the interaction of studied binding partners.

[0151] The two-hybrid approach was first developed for yeast, and is anideal screening system when looking for compounds active in killingyeast or fungi. Indeed, proteins expressed in this system will mostprobably carry the correct modifications as found in the pathogenicyeast strains. In addition, compounds active in this test system allowto screen and select compounds which are able to enter the cell, thisselection is not possible when using in vitro test systems. Whencompounds are needed to target mammalian cells, modification of thestudied proteins can be different, changing the structure ofcorresponding proteins. Moreover working with yeast might block certaincompounds to enter the cell, which are normally able to traverse themammalian cell membrane. Consequently, working with mammalian two-hybridsystem for this purpose will give already an immediate selection of thecompounds that may enter mammalian cells.

[0152] Alternative in vitro methods can be used to investigateprotein-protein interactions. Protein interaction analysis in vitro canshed light on their role in the intact cell by providing valuableinformation on specificity, affinity, and structure-function relationship. Significant progress in this respect has become with the advent,in the last few years, of commercially available biosensor technology.This allows to study macromolecular interactions in real-time, providinga wealth of high-quality data that can be used for kinetic analysis,affinity measurements, competition studies, etc. A major advantage ofbiosensor analysis is that there is no requirement for labeling one ofthe interacting components and then separating bound from freemolecules—a fact that simplifies experimental procedures and providesmore accurate measurements. The principle of surface plasmon resonance(SPR) is based on the detection of a change of the refractive index ofthe medium when a compound or protein binds to an immobilised partnermolecule. For the SPR technology, one needs to load one of theinteracting partners to the chip surface, followed by the superfusion ofthe second binding partner or more molecules. The second partner can beavailable as purified product, but alternatively a complex suspensioncontaining this partner can also be used. Interaction of two or morecompounds can be analysed, alternatively, compounds can be identifiedinterfering or increasing this binding affinity towards each other.

[0153] SPR is not restricted to protein-protein interactions; anymacromolecule with a suitable size will change the refractive index ofthe medium in contact with the biosensor surface and therefore give asignal. Studies have been done with protein-DNA interactions, as well asprotein-lipid interactions. Moreover intact viruses, and even cells, canalso be injected over the biosensor surface, in order to analyse theirbinding to receptors, lectins, and so on.

[0154] Alternatively, NMR is also an excellent tool for a detailed studyof protein-protein or DNA-protein interactions. Isotope edited orisotope filtered experiments whereby one compound is isotopicallylabeled with ¹⁵N or ¹³C are an ideal way to study these complexes. Thismethod does not allow high throughput analysis of compounds interferingor enhancing molecular interactions. Nevertheless, medium or lowthroughput systems can be used to confirm results obtained by the highthroughout assays or in cases where none of the binding partners arelabeled. Other techniques which can be used to study interactions are:overlay, ligand blotting, band-shift, co-immuno-precipitation, sizeexclusion chromatography and microcalorimetry (In. “Protein targetingProtocols” Ed. Clegg R. A. Humana Press, Totowa, New Yersey).

[0155] Compounds modulating pathways leading to apoptosis may change theactivity of the polypeptide of the invention. Therefore screening testsmay be setup looking for altered protein activity of the polypeptide ofthe invention. Based on the amino acid sequence a possible function ofthe polypeptide might be envisaged; activities can be confirmed andcorresponding activity test can be started.

[0156] Alternatively additional tests can be performed to test theinfluence of the compound onto protein stability, post-translationalmodification, precursor processing and protein translocation. All theseaspects influence the concentration and/or activity of correspondingproteins and consequently influence the effect of these onto themetabolism of the cell. Also here, medium or low throughput systems canbe used to confirm results obtained by the high throughout assays.

[0157] In cases compounds need to be found to target tumor cells,screening assays will have to be used focused on the stimulation of theapoptotic pathway. This invention therefore also relates to in vitro andin vivo model systems comprising tumor tissue or cells expressing thepolypeptides according to the invention which can be used to screen fortherapeutic agents. In vivo modelsystems allow to test for compoundefficacity but also the toxicity of these compounds can be tested. Thecompounds identified using any of the methods described in the inventionnot only include compounds which exert their effect in promoting celldeath of yeast and fungi, but also include compounds which prevent ordelay cell death. The latter compounds can be used to prevent or delayapoptosis of endogenic yeast or fungi in humans and other mammals whichmay be caused by pathogens or toxic environmental components.

[0158] According to a preferred aspect of the invention, the yeast orfungi according to any of the methods described, are chosen from Candidaspp., Aspergillus spp., Microsporum spp., Trichophyton spp., Fusariumspp., Zygomycetes spp., Botritis, spp., Cladosporium spp., Malasseziaspp., Epidermophyton floccosum, Blastomyces dermatitidis, Coccidioidesimminitis, Histoplasma capsulatum, Paracoccidioides brasiliensis,Cryptococcus neoformans, and Sporothrix schenckii.

[0159] The invention also relates to a compound identified using any ofthe methods of the invention. Compounds identifiable or identified usinga method according to the invention, may advantageously be used as amedicament. The invention also relates to a method for treating diseasesassociated with yeast or fungi comprising admixing a compound obtainableby a method of the invention with a suitable pharmaceutically acceptablecarrier.

[0160] The invention further relates to a method for preparingpharmaceutical composition for treating diseases associated with yeastor fungi comprising admixing a compound as identified above with asuitable pharmaceutically acceptable carrier. The invention also relatesto said pharmaceutical composition.

[0161] The compounds or pharmaceutical compositions of the invention canbe used for the preparation of a medicament to treat diseases orconditions associated with yeast and fungi infections, more preferablywhere the yeast or fungus is chosen from Candida spp., Aspergillus spp.,Microsporum spp., Trichophyton spp., Fusarium spp., Zygomycetes spp.,Botritis, spp., Cladosporium spp., Malassezia spp., Epidermophytonfloccosum, Blastomyces dermatitidis, Coccidioides imminitis, Histoplasmacapsulatum, Paracoccidioides brasiliensis, Cryptococcus neoformans, andSporothrix schenckii.

[0162] These compounds may also advantageously be included in apharmaceutical composition together with a pharmaceutically acceptablecarrier, diluent or excipient therefor.

[0163] A medicament according to the invention not only relates tofungicidal and fungistatic compounds for treating humans or mammals butalso relates to fungicides for treating plants.

[0164] According to yet another embodiment, the invention relates to agenetically modified yeast or fungus in which modification results inthe overexpression or underexpression of at least one of the nucleicacids or the polypeptides of the invention, which overexpression orunderexpression of said nucleic acid or polypeptide prevents, delays orsensitizes for apoptosis of said genetically modified yeast or fungus.These genetically modified organisms may have a positive effect on theendogenic flora of humans and other mammals. The genetically modifiedyeast or fungi can be included in a pharmaceutical composition or can beused for the preparation of a medicament for prophylactic or therapeuticuse.

[0165] Also according to the invention is the use of a compoundobtainable by a method of the invention, a pharmaceutical composition ora genetically modified organism as described above for the preparationof a medicament for modifying the endogenic flora of humans and othermammals.

[0166] According to another embodiment, the invention relates to agenetically modified mammalian cell or non-human organism in whichmodification results in the overexpression or underexpression of atleast one of the nucleic acids of the invention or a human homologuethereof or at least one of the polypeptides of the invention or a humanhomologue thereof, which overexpression or underexpression of saidnucleic acid or polypeptide prevents or delays apoptosis of saidgenetically modified mammalian cell or in said genetically modifiednon-human organism.

[0167] According to a preferred embodiment, the invention relates to agenetically modified mammalian cell or non-human organism as describedabove wherein said modification comprises the expression of an antisensemolecule to at least one of the nucleic acids of the invention or anantisense molecule to a mammalian homologue of said nucleic acid.

[0168] The invention also relates to a method for identifying compoundsfor stimulating or inhibiting apoptosis comprising the use of at leastone of the nucleic acid sequences of the invention or a human homologuethereof and/or at least one of the polypeptides of the invention or ahuman homologue thereof and/or a genetically modified mammalian cell ornon-human organism as described in the invention.

[0169] Some examples of preferred human homologues of yeast and/orCandida spp. sequences which can be used in the above methods arerepresented in SEQ ID NOs 675 to 686.

[0170] The invention further relates to the compounds identifiableaccording to the above-described method and their use as a medicament.

[0171] The invention further relates to a method for preparing apharmaceutical composition for treating proliferative disorders or forpreventing apoptosis in certain diseases comprising admixing a compoundidentifiable according to the above-described methods with a suitablepharmaceutically acceptable carrier.

[0172] The invention also relates to the use of compounds obtainable bythe above described methods for the preparation of a medicament fortreating proliferative disorders or for preventing apoptosis in certaindisorders.

[0173] Furthermore, the present inventors overexpressed the Bax proteinin the pathogenic yeast Candida albicans and found that this leads to asimilar phenotype. However these results could only be received afterhaving constructed a new synthetic bax gene which could be adequatelyexpressed in this pathogenic organism.

[0174] Therefore, the present invention relates to an isolated nucleicacid representing a synthetic BAX-gene for expression in Candida spp.selected from the group of:

[0175] a) a nucleic acid comprising a sequence as represented by SEQ IDNO 1,

[0176] b) a nucleic acid comprising a fragment of a sequence of SEQ IDNO 1 and encoding a functional fragment of the sequence represented bySEQ ID NO 2,

[0177] c) a nucleic acid comprising a sequence as represented in any ofSEQ ID NOs 3 to 10,

[0178] d) a nucleic acid which is more than 75% identical, preferablymore than 80%, 85%, 90% or 95% identical, most preferably more than 97%identical to the nucleic acid as represented by SEQ ID NO 1, or to anucleic acid according to the nucleic acid as defined in b) or c), and

[0179] e) a nucleic acid as defined in any one of (a) to (i) interruptedby intervening DNA sequences,

[0180] or a nucleic acid representing the complement of any of saidnucleic acids as defined in (a) to (d).

[0181] The synthetic BAX gene shows 73.7% identity with the gene codingfor Bax-a. It should be clear that the present invention also relates tonucleic acids wherein other, also frequently used Candida spp. codons,are used instead of the choice made for the sequence represented in SEQID NO 1. (Table 8)

[0182] It should be clear that all nucleic acids according to theinvention and which are specifically described above, can be DNA, cDNA,genomic DNA, synthetic DNA, or RNA wherein T is replaced by U.

[0183] According to another embodiment of the invention, the nucleicacid sequences according to the invention as defined above may,advantageously, be included in a suitable vector, preferably anexpression vector which may be transformed, transfected or infected intoa host cell. In such an expression vector the nucleic acid is operablylinked to one or more control sequences allowing the expression in hostcells, such as a suitable promotor, or the like, to ensure expression ofthe proteins according to the invention in a suitable prokaryotic oreukaryotic host cell. In this respect, a constitutive or an induciblepromoter can be used.

[0184] As described in the examples, the invention also relates tonucleic acids and constructs comprising the synthetic BAX, or partsthereof, as a fusion with a carrier gene, such as, but not restricted tothe yeast GFP gene. It is not necessary to include the complete gene ofthe fusion partner in the expression construct, so the invention relatesto various fusion products which can result from the synthetic BAX geneand its fusion partner.

[0185] The expression vectors comprising the synthetic construct orfusion protein and the host cell defined herein also form part of thepresent invention. Said host cell can be from bacterial, yeast, fungal,insect, mammal or human origin. An interesting host cell according tothe invention is a Candida spp. cell.

[0186] In another embodiment, the expression vector may further comprisean inducible promoter, and/or further a reporter molecule.

[0187] The invention also relates to a vector as described above forinducing programmed cell death in Candida spp.

[0188] The invention further also relates a genetically modified yeastor fungal cell as described above wherein said modification results inthe onset of at least one pathway eventually leading to programmed celldeath.

[0189] The invention also relates to a genetically modified Candida spp.cell wherein said modification results in the onset of at least onepathway eventually leading to programmed cell death

[0190] According to a further embodiment, the invention relates to amethod for identifying genes in Candida spp. which are differentiallyexpressed in a pathway eventually leading to programmed cell death usinga synthetic BAX gene, as described above, or a vector comprising saidgene as described herein, or a genetically modified yeast or fungal cellas described above.

[0191] In this respect different model systems are envisaged. It hasbeen shown in the present invention that expression of the synthetic BAXgene as a fusion protein more rapidly kills the host cells than whenexpressed without a fusion partner. Accordingly there will be adifference in which Candida spp. genes will be differentially expressedin each system. The invention thus relates to methods for identifyinggenes in Candida spp. which are differentially expressed in a pathwayeventually leading to programmed cell death, wherein in said methods thehost cells will need a longer or shorter time period for starving. Saidtime period is dependent on the expression construct or system used.

[0192] The invention further relates to a method for obtaining andidentifying Candida spp. sequences (genes or polypeptides) involved in apathway eventually leading to programmed cell death comprising the stepsof:

[0193] a) providing a two hybrid system wherein a polypeptide encoded bya nucleic acid as described above or a vector as described above as abait and a Candida spp. cDNA library as a prey are expressed,

[0194] b) detecting an interaction between said polypeptide and aCandida spp. polypeptide encoded by said cDNA library, and,

[0195] c) identifying said Candida spp. polypeptide.

[0196] The invention also relates to a method for identifying inhibitors(or inhibitor sequences) of Bax-induced cell death comprising the stepsof:

[0197] a) providing a genetically modified organism as described above,

[0198] b) expressing a cDNA library in said genetically modifiedorganism, and,

[0199] c) identifying a polypeptide or a cDNA which expression has abeneficial effect on the survival and/or growth of said geneticallymodified organism.

[0200] The invention further relates to a method for identifyingBax-resistant yeast or fungi comprising the steps of:

[0201] a) providing (a) genetically modified yeast or fungi as describedabove,

[0202] b) treating said genetically modified yeast or fungi with amutagen,

[0203] c) isolating resistant yeast or fungal cells, and,

[0204] d) optionally identifying and/or characterizing mutated genes insaid resistant yeast or fungal cells.

[0205] The invention further relates to any of the methods describedabove wherein said genetically modified organism is a Candida spp.

[0206] The invention also relates to an isolated Candida spp. nucleicacid identifiable by any of the methods described above.

[0207] The invention, now being generally described, may be more clearlyunderstood by reference to the following examples, which are includedmerely for purposes of illustration of certain aspects and embodimentsof the present invention and are not intended to limit the invention.The contents of all references referred to in this text are herebyincorporated by reference.

FIGURE AND TABLE LEGENDS

[0208]FIG. 1. Saccharomyces cerevisiae sequences based on informationobtained from the Saccharomyces Genome Database (SGD) (SEQ ID NOs 17 to396 and SEQ ID NOs 691 to 716)

[0209]FIG. 2. Candida albicans (SEQ ID NOs 397 to 674, 687, 688 and 717to 732) and human homologues (SEQ ID NOs 675 to 686).

[0210] Human homologues were confirmed via forward and reverse BLASTusing BLOSUM62 as a scoring matrix.

[0211] YGL080W (SEQ ID NO 161) codes for a yeast protein with an unknowncellular role and an unknown biochemical function. The human homologue(330 bp (SEQ ID NO 675), 109 aa (SEQ ID NO 676)) LOC51660/g7706369 hasno reported cellular role or biochemical function.

[0212] YGR243W (SEQ ID NO 189) codes for a yeast protein with an unknowncellular role and an unknown biochemical function. The human homologue(384 bp (SEQ ID NO 677), 127 aa (SEQ ID NO 678)) DKFZP564B167/g5817257has no reported cellular role or biochemical function.

[0213] YGR183C (QCR9) (Table 3) codes for a yeast protein with a knowncellular role and a known biochemical function. QCR9 codes for subunit 9of ubiquinol cytochrome-c reductase (7.3 kDa protein) which is acomponent of the ubiquinol cytochrome-c reductase complex. Cellularrole: energy generation. Biochemical function: oxidoreductase and activetransporter. The human homologue (132aa (SEQ ID NO 679), 399 bp (SEQ IDNO 680)) AF161536 was predicted to have an analogous cellular role andbiochemical function.

[0214] YBR009C (SEQ ID NO 37), YGR209C (SEQ ID NO 187) and YPR028W (SEQID NO 393) correspond to known yeast ORFs. Their human homologues have areported cellular role or biochemical function.

[0215]FIG. 3. Yeast genome macroarray containing a total of 6144 geneORFs spotted on 2 nylon membrane filters (I and II). Each filtercontains 2 fields and each field is divided into 8 grids, organised in24 rows and 8 columns.

[0216] The spots represent the genome wide expression profile without(Minus BAX) and with (Plus BAX) induction of Bax expression for 30 min,1 hour, 2 hours, 3 hours and 6 hours.

[0217]FIG. 4 Yeast cells with a disrupted YGR183C gene are fullyresistant to Bax-induced cell death. Resistance is observed in both thelow-copy (A) and the high-copy (B) Bax expression system. Clonogenicsurvival was determined by recovering cells at various times fromgalactose-containing medium and plating of 1000 cells on glucose-basedsemisolid medium. Data are representative of three experiments (mean±SD,n=3). SD bars are obscured by symbols.

[0218]FIG. 5. Scheme for the synthesis of the synthetic BAX gene usingC. albicans optimal codons.

[0219]FIG. 6. DNA (SEQ ID NO 1) and protein (SEQ ID NO 2) sequence ofthe synthetic C. albicans BAX gene.

[0220]FIG. 7. Representation of the expression constructs of thesynthetic CaBAX gene (A) and the yEGFP-synth CaBAX fusion (B).

[0221]FIG. 8. Growth of the Candida Albicans transformants: theindividual transformants of pGAL1P:synthCaBAX and pGAL1P:GFP-synthCaBAXwere streaked onto plates containing either 2% glucose or 2% galactoseas sole carbon source. Growth was monitored 4 days later.

[0222]FIG. 9. Growth kinetics of GALL P:synthCaBAX (A) and GALLP:GFP-synthCaBAX (B) on galactose containing minimal medium.

[0223]FIG. 10. Immunoblot analysis of two independent transformants ofGAL1P:synthCaBAX after 15 hours Bax induction on minimal galactosecontaining media. The arrow at 20 kDa indicates the position of the Baxprotein. The band seen at 50 kDa probably represents a cell wall mannan.Not all of the contamination of the polyclonal Bax antibody could beremoved by the threatment with S. cerevisiae mannan.

[0224]FIG. 11. Immunoblot analysis of the GAL1P:GFP-synthCaBAX strain ongalactose containing minimal medium. The band appearing at 45 kDarepresents the Gfp-Bax fusion protein, while the band at 20 kDarepresents the Gfp protein alone.

[0225]FIG. 12. FACS analysis of two independent GAL1P:GFP-synthCaBAXtransformants grown on galactose containing media: the light grey peakindicates the autofluorescence of the wt strain, the GFP-fluorescencepeak is not shaded.

[0226]FIG. 13. Viability test synthCaBAX (A) and GFP-synthCaBAXtransformants (B): Cells were pregrown in minimal dextrose medium andthen switched to fresh minimal medium containing galactose. At the timepoints indicated, samples were taken and equal cell amounts were spreadon minimal dextrose plates. The appearing colonies represented theviable fraction of the total pool.

[0227] Table 1. Oligonucleotides used for construction of the syntheticCaBAXx gene: start and stop codon are in bold, restriction sites usedfor cloning are in bold and italic.

[0228] Tables 2-6. Genes modulated by Bax expression in S. cerevisiae.This list includes the genes for which mRNA levels changed significantlyafter a 30 min (Table 2), 1 hour (Table 3), 2 hours (Table 4), 3 hours(Table 5) or 6 hours (Table 6) induction of Bax protein expression. TheQt values were calculated using the Pathways™ software (ResearchGenetics).

[0229] Table 7. Genes modulated by Bax expression in S. cerevisiae. Thislist includes all the genes for which mRNA levels changed significantlyafter induction of Bax protein exppression. The Ot values werecalculated using the Pathways software (Research Genetics). Positivevalues correspond with upregulated genes. Negative values correspondwith downregulated genes. (Comparable with ↑ and ↓ respectively inTables 2-6).

[0230] Table 8. Codon usage for the synthetic BAX gene.

[0231] Table 9. Regulation of 23 selected “Bax-specific” functions.

EXAMPLES Example 1 Differential Gene Expression Analysis UponBax-Induced Cell Death

[0232] Materials and Media

[0233] Bacterial strain Escherichia coli MC1061 (Casadaban and Cohen,1980) was used for the construction and the amplification of plasmids.Yeast strains were grown under normal conditions on standard media(Sherman et al., 1979). The Saccharomyces cereviseae strain INVSc1(Invitrogen®, San Diego, Calif., USA) was transformed by means of thelithium acetate method (Schiestl and Gietz, 1989) with YIpUTyL orYIpUTYLMuBax, after linearisation in the Ty δ element (Zhu, 1986).

[0234] Cloning of Mouse BAX cDNA

[0235] Mouse bax cDNA, encoding the mouse Bax-α protein, was cloned byPfu DNA polymerase (Stratagene®, Lo Jolla, Calif., USA) chain reactionamplification (PCR) from an EL4/13.18 thymoma cDNA library(BCCM™/LMBP-LIB15) by making use of the primers:5′-ATGGACGGGTCCGGGAGCAG-3′ (SEQ ID NO 689) and5′-TCAGCCCATCTTCTTCCAGATGGTGAG-3′. (SEQ ID NO 690)

[0236] The resulting PCR product was cloned in a HincII-openend pUC19according to standard procedures (Sambrook J. et al., 1989).

[0237] Plasmid Constructions

[0238] The 2μ ori and the URA3 marker gene were removed from pUT332(Gatignol et al., 1990) by successive digestions with ClaI and BglII. ABamHI-HindIII GAL1 promoter fragment was ligated into theBglII-HindIII-opened plasmid. A XbaI-FspI FLP terminator fragment wasinserted into this XbaI-HindIII(blunted)-opened plasmid so that theplasmid YIpUT was obtained. Insertion of a blunted EcoRI-BsaAI Ty δelement in the KpnI-AatII-opened and blunted YIpUT resulted in theplasmid YIpUTy. Subsequent insertion of the LEU2 marker gene, as ablunted BsaAI-BsrGI fragment, in the BamHI-openend and blunted YIpUTyresulted in the plasmid YIpUTyL.

[0239] Mouse bax cDNA was excised from pUC19 by digestion with XbaI andHindIII and subcloned into the XbaI-HindIII-opened plasmid YIpUTyL,obtaining the final expression plasmid YIpUTyLMuBax.

[0240] The plasmid YIpUTyLMuBax has been deposited in the BCCM™/LMBPculture collection as pSCTyGALmBax with accession number 3871 underrestricted use.

[0241] GeneFilters

[0242] The Yeast GeneFilters™ were purchased from Research Genetics Inc.(Huntsville, Ala., USA).

[0243] The Yeast GeneFilters™ are hybridization ready nylon membranescontaining a total of 6144 gene ORFs (Open Reading Frames) individuallyamplified by PCR and spotted on 2 nylon membrane filters (Filter I andII). The filters are cut in the upper right corner and the DNA is on thelabeled side of the filter.

[0244] Filter I contains 3072 ORFs organized into two fields (fields 1and 2). Each field contains 1536 ORFs divided into 8 grids (A, B, C, D,E, F, G and H). The grids are organized in 24 rows and 8 columns.

[0245] Filter II contains 3072 ORFs organized in two fields (field 3 and4). Fields 3 and 4 are organized in the same way as fields 1 and 2.

[0246] The Yeast ORF Target

[0247] The yeast filters consist of over 6144 PCR products correspondingto 6144 yeast ORFs derived from the SGD. The PCR reactions used ORFspecific primer pairs designed to amplify the entire open reading frame.The primers were generated from unique sequences containing the startcodon ATG and termination codon (kindly provided by M. Cherry atStanford Genome Center). Thus the PCR product contains the complete openreading frame including the start and stop codons. These products werepurified and resuspended at 50 nanograms per microliter in a coloredsolution to allow the printing to be monitored. A robotic device wasused to spot approximately 1/10 of a microliter of the denatured PCRproduct solution on a positively charged nylon membrane. The DNA wasthen UV cross-linked to the membrane.

[0248] Results

[0249] Induction of Bax-Expression in Yeast Cells

[0250] A preculture of yeast strain INVSc1 containing YIpUTyLMuBax,wherein 5 Bax cassettes under the control of the GAL1 promotor areintegrated in the genome near Ty δ elements, was grown overnight inminimal glucose-containing medium in parallel with the yeast strainINVSc1 containing YIpUTyL as a control. The precultures were diluted in100-ml minimal glucose-containing medium and grown until an OD₆₀₀ of 1was reached. Subsequently, the yeast cells were transferred into 100-mlgalactose-containing medium and incubated for an additional period of 30min, 1 hour, 2 hours, 3 hours or 6 hours.

[0251] RNA Isolation

[0252] Total RNA was isolated using RNApure™ Reagent (GenhunterCorporation Nashville, Tenn., USA) according to the GenHunter protocol.1.5 10⁹ cells were concentrated in a microcentrifuge tube and 1 mlRNApure™ Reagent was added together with 1 g of glass pearls. The yeastcells were broken by thorough mixing during five 2-minutes periods, andplaced on ice in-between to avoid RNA degradation. Chloroform (150 μl)was added to the lysate and centrifuged for 10 min at 4° C. and at 15000rpm. The supernatant was transferred to a new tube and the RNA wasprecipitated with an equal volume of isopropanol. After 10 minincubation on ice, the RNA was pelleted by centrifugation and the pelletwas washed with 70% ice-cold ethanol. The dried RNA pellet wasresuspended in 50 pi RNAse free dH₂O.

[0253] First Strand cDNA Synthesis in the Presence of α-³³P dCTP

[0254] Probes with high specific activity were prepared by first strandcDNA synthesis using total RNA isolated from INVSc1 YIpUTyLMuBax orINVSc1 YIpUTyL yeast cells and incorporation of α-³³P dCTP as follows: 2μl (1 μg/ml) of Oligo dT was added to 20 μg of total RNA in a maximalvolume of 8 μl RNase-free dH₂O and incubated at 70° C. for 10 min. Aftercooling down on ice for 1 min, the following components were added:

[0255] 6 μl 5× concentrated First Strand Buffer (GIBCO-BRL, Paisley, UK)

[0256] 1 μl 0.1 M DTT

[0257] 1 μl RNase Block (40 units/μl) (Stratagene)

[0258] 1.5 μl 20 mM dXTP-solution (X=A, G and T) (Amersham Pharmaciabiotech Uppsala, Sweden)

[0259] 1.5 μl SuperScript™ Reverse Transcriptase (200 units/μl)(GIBCO-BRL)

[0260] 10 μl α-³³ P dCTP (10 mCi/ml, 3000 Ci/mmol) (Amersham Pharmaciabiotech Uppsala, Sweden),

[0261] and incubated for 2 h at 37° C. during which first strand cDNAsynthesis took place.

[0262] Unincorporated label was separated from the probe on a SephadexG-50 column (Amersham Pharmacia biotech Uppsala, Sweden). Theradioactivity incorporated in the probe was measured by liquidscintillation. The specific activity of the probes was 5.10⁸ cpm/μg forboth the INVSc1YIpUTyL and the INVSc1 YIpUTyLMuBax probes.

[0263] Additionally, the length of first strand cDNA probes wascontrolled on an alkaline 2% agarose gel using standard electrophoresistechniques, and resulted in the detection, via stimulatedphosphorescence autoradiography, of the bulk of the fragments around 500bp.

[0264] Hybridisation with the S. cerevisiae Yeast GeneFilters™ andSignal Detection

[0265] The Yeast GeneFilters™ were successively hybridised with theα-³³P dCTP labelled cDNA probes using the MicroHyb™ solution provided bythe manufacturer (Research Genetics Inc., Huntsville, Ala., USA). Thissolution was applied as well in the prehybridisation step as duringhybridisation. The MicroHyb™ solution contains formamide to allowhybridisation to occur at lower temperatures.

[0266] The hybridisation experiment was performed essentially asfollows: during prehybridisation, the Yeast GeneFilters™ were placed ina hybridisation flask (35×250 mm) filled with 5 ml MicroHyb™ solution(42° C.) containing 5 μl polydA (1 μg/ml) and incubated for 24 hours at42° C. whilst rotating (10 rpm). After disposal of the prehybridisationsolution, the denatured (3 min at 100° C.) cDNA was added in 5 mlprewarmed MicroHyb solution and again incubated overnight at 42° C.whilst rotating. Following two wash steps of 20 min in wash buffer(2×SSC, 1% SDS) at 50° C., a third wash step was performed in a secondwash buffer (0.5×SSC, 1% SDS) for an additional 15 min at roomtemperature. The Yeast GeneFilters™ were placed in a Phosphorlmager™cassette (Molecular Dynamics, Sunnyvale, Calif., USA) with storagephosphor screen. After 4 days of development the screen was scanned at aresolution of 50 μm using the (BioRad, Richmond, Calif., USA) PersonalFX. The results of these can be seen in FIG. 3.

Example 2 Quantification of Hybridisation Signals

[0267] Quantification of the hybridisation signals was done using thePathways™ software (Research Genetics, Huntsville, Ala., USA) and thesesignals were normalised against all data points. Comparison of thesenormalised data revealed differentially expressed candidate genes.Visual inspection of the hybridisation spots confirmed their selection.The genes as well as the factors with which they are up- ordown-regulated are listed in the Tables 2 to 6 for each individual timepoint. An overview of the up and down regulated genes modulated infunction of induction of Bax expression for several time points is shownin Table 7. The sequences of these genes and amino acid sequences thatthey encode are shown in FIG. 1.

Example 3 Comparative Gene Expression Analysis Upon Bax-Induced CellDeath and H₂O₂-Induced Cell Death

[0268] The Oxidative H₂O₂-Challenge

[0269] A preculture of yeast strain INVSc1 containing YIpUTyL was grownovernight in minimal glucose-containing medium. The preculture wasdiluted in 100-ml minimal glucose-containing medium and grown until anOD₆₀₀ of 1 was reached. Subsequently, the yeast cells were transferredinto 100-ml galactose-containing medium supplemented with 0.1 mM H₂O₂,and incubated for an additional period of 1 hour. This oxidativechallenge resulted in the same final toxicity as a 1-hour induction ofBax expression in the same growth conditions.

[0270] First Strand cDNA Synthesis in the Presence of α-³³P dCTP

[0271] RNA was isolated as mentioned in Example 1. Probes with highspecific activity were prepared (detailed in Example 1) by first strandcDNA synthesis using total RNA isolated from INVSc1 YIpUTyLMuBax orINVSc1 YIpUTyL (growth conditions as described in Example 1) oroxidatively stressed INVSc1 YIpUTyL yeast cells.

[0272] The specific activity of all probes was 5.10⁸ cpm/μg.

[0273] Quantification of Hybridisation Signals

[0274] Hybridisation and signal detection as described in Example 1.Conversion of the digital images to a 16 bit TIFF format using theQuantity One program (BioRad, Hercules, Calif., USA) preserved imagedata and was necessary for file import into the Pathways® software(Research Genetics, Huntsville, Ala., USA). Pathways® was used for thequantification of hybridisation signals and these signals werenormalised against all data points.

[0275] Identification of Bax-Responsive Genes

[0276] Pairwise comparisons of the normalised data obtained from INVSc1YIpUTyLMuBax (B) and INVSc1 YIpUTyL (C) revealed differentiallyexpressed genes. To determine the -fold induction or repression, thenormalised signal intensity after Bax induction (B) was divided by thatbefore the shock (C). Visual inspection of the hybridisation spotsconfirmed their selection (replacement).

[0277] Identification of Bax-Specific Genes within the Bax-ResponsivePool

[0278] Pairwise comparisons of the normalised data obtained from INVSc1YIpUTyLMuBax (B) and INVSc1 YIpUTyL (C) at the 1-hour time pointrevealed differentially expressed genes. Linear ratios (B vs C) wereestimated significant when changes were at least two-fold and thenormalised signal intensity of one spot was at least tenfold above theaverage background value. The normalised data of the Bax-responsivegenes were compared with data obtained from the H₂O₂-stressed INVSc1YIpUTyL (H). A Bax-responsive (up-regulated/down-regulated) gene wasconsidered to be Bax-specific when the normalised signal intensity afterBax induction was at least twice as high/low as the correspondingintensity after oxidative stress. Visual inspection of the hybridisationspots confirmed their selection. An overview of the Bax-specific genesfor the 1-hour time point is shown in Table 9. The sequences of thesegenes and amino acid sequences that they encode are shown in FIG. 2.

Example 4 Search for Homologues in Candida albicans and Human

[0279] Sequence similarity searches against public and commercialsequence databases were performed with the BLAST software package(Altschul et al., 1990) version 2. Both the original nucleotide sequenceand the six-frame conceptual translations were used as query sequences.The used public databases were the EMBL nucleotide sequence database(Stoesser et al., 1998), the SWISS-PROT protein sequence database andits supplement TrEMBL (Bairoch and Apweiler, 1998), and the ALCESCandida albicans sequence database (Stanford University, University ofMinesota). The commercial sequence database used was the PathoSeq™microbial genomic database (Incyte Pharmaceuticals Inc., Palo Alto,Calif., USA).

[0280] Sequence similarity searches were performed using the BLASTsoftware package version 2. The identity between 2 sequences wascalculated as percentage identical residues, the similarity percentagebetween two sequences was calculated using BLOSUM62 as a scoring matrix.

[0281] The sequences of homologues Candida spp. and human genes and thecorresponding amino acid sequences are shown in FIG. 2.

Example 5 Screening for Compounds Modulating Expression of PolypeptidesInvolved in Induction of Cell Death of C. albicans

[0282] The method proposed is based on observations (Sandbaken et at,1990; Hinnebusch and Liebman 1991; Ribogene PCT WO 95/11969, 1995)suggesting that underexpression or overexpression of any component of aprocess (e.g. translation) could lead to altered sensitivity to aninhibitor of a relevant step in that process. Such an inhibitor shouldbe more potent against a cell limited by a deficiency in themacromolecule catalyzing that step and/or less potent macromolecule, ascompared to the wild type (WT) cell.

[0283] Mutant yeast strains, for example, have shown that some steps oftranslation are sensitive to the stoichiometry of macromoleculesinvolved. (Sandbaken et al, 1990). Such strains are more sensitive tocompounds which specifically perturb translation (by acting on acomponent that participates in translation) but are equally sensitive tocompounds with other mechanisms of action.

[0284] This method thus not only provides a means to identify whether atest compound perturbs a certain process but also an indication of thesite at which it exerts its effect. The component which is present inaltered form or amount in a cell whose growth is affected by a testcompound is potentially the site of action of the test compound.

[0285] The assay to be set up involves measurement of growth and/ordeath rate of an isogenic strain which has been modified only in acertain specific allele, relative to a wild type (WT) Candida albicansstrain, in the presence of R-compounds. Strains can be ones in which theexpression of a specific protein is impaired upon induction ofanti-sense or strains which carry disruptions in an essential gene. Anin silico approach to find novel genes in Candida albicans will beperformed. A number of essential genes identified in this way will bedisrupted (in one allele) and the resulting strains can be used forcomparative growth and/or death rate screening.

Example 6 Assay for High Throughput Screening for Drugs

[0286] 35 μl minimal medium (S medium+2% galactose+2% maltose) istransferred in a transparent flat-bottomed 96 well plate (MW96) using anautomated pipetting system (Multidrop, Labsystems, Helsinki, Finland). A96-channel pipettor transfers 2.5 μi of R-compound at 10⁻³ M in DMSOfrom a stock plate into the assay plate.

[0287] The selected Candida albicans strains (mutant and parent (CAI-4)strain) are stored as glycerol stocks (15%) at −70° C. The strains arestreaked out on selective plates (SD medium) and incubated for two daysat 30° C. For the parent strain, CAI-4, the medium is alwayssupplemented with 20 μg/ml uridine. A single colony is scooped up andresuspended in 1 ml minimal medium (S medium+2% galactose+2% maltose).Cells are incubated at 30° C. for 8 hours while shaking at 250 rpm. A 10ml culture is inoculated at 250.000 cells/ml. Cultures are incubated at30° C. for 24 hours while shaking at 250 rpm. Cells are counted inCoulter counter and the final culture (S medium+2% galactose+2% maltose)is inoculated at 20.000 to 50.000 cells/ml. Cultures are grown at 30° C.while shaking at 250 rpm until a final OD₆₀₀ of 0.24 (+/−0.04) isreached.

[0288] 200 μl of this yeast suspension is added to all wells of MW96plates containing R-compounds in a 450 p, total volume. MW96 plates areincubated (static) at 30° C. for 48 hours.

[0289] Optical densities are measured after 48 hours.

[0290] Test growth is expressed as a percentage of positive controlgrowth for both mutant (x) and wild type (y) strains. The ratio (x/y) ofthese derived variables is calculated.

Example 7 Yeast Cell Viability Assay Upon Induction of Bax Expression

[0291] Materials and Media

[0292] Yeast stains were grown under normal conditions on standard media(Sherman et al., 1979). The Saccharomyces cerevisiae BY4742 wild typestrain and BY4742 with the YGR183C gene disruption (EUROSCARFcollection) were transformed by means of the lithium acetate method(Schiestl and Gietz, 1989) with the low-copy centromeric pRS415Baxplasmid or pRS415 as a control, or with the high-copy episomal pRS425Baxplasmid or pRS425 as a control.

[0293] Plasmid Constructions

[0294] The Bax expression cassette, a BsgI(blunted)-SapI(blunted)fragment excised from YIpUTyLMuBax containing the GALL promoter, the baxcDNA and the FLP terminator, was ligated into the Ectl3611-opened pRS415(ATCC 87520) and pRS425 (ATCC 77106) plasmids, obtaining the low-copycentromeric pRS415Bax and the high-copy episomal pRS425Bax expressionplasmids.

[0295] Results

[0296] Single colonies of yeast cells transformed with pRS415 orpRS415Bax or pRS425 or pRS425Bax were grown in 10 ml minimalglucose-containing medium with vigorous aeration at 30° C. to an opticaldensity of 1 OD₆₀₀. Cells were pelleted by centrifugation and washed twotimes with sterile dH₂O before resuspending in 10 ml minimalgalactose-containing medium. After culturing for various times at 30°C., the total cell density of the cultures was determined, and 1000cells were spread on minimal glucose-based semisolid medium, followed byincubation at 30° C. for 3 days. The number of colonies on plates fromthe 0 hr cultures was designated as 100% (FIG. 4).

Example 8 Bax Expression in Candida Cells

[0297] Strains

[0298] The Candida albicans strain CAI4 (ura3≅) was used to perform theexperiments (Fonzi and Irwin 1993).

[0299]E. coli transformations were done using the Top10 strain fromInvitrogen (San Diego, Calif., USA) (F′ mcrA ≅(mrr-hsdRMS-mcrBC)≅80lacZ□M15 ≅lacX74 deoR recA1 araD139 ≅(ara/leu)7697 galU galK rpsL(Str^(R)) endA1 nupG).

[0300] Media

[0301] Synthetic dextrose media (SD), containing 2% glucose, 1.34% YeastNitrogen Base without amino acids and 0.77 g/l CSM-ura (Bio 101, Vista,Calif., USA) was used to grow the Candida albicans transformants. Incase of the wild type (CAI4), the media was supplemented with 50 μg/mluridine. To prepare plates the media was solidified with 2% agar.Expression of the synthetic BAX gene was performed using 2% galactose ascarbon source.

[0302] Construction of the Codon-Optimised BAX Gene

[0303] Construction of the synthetic BAX gene followed the nomenclaturedescribed for Candida albicans (Lloyd and Sharp 1992; Brown, et al.1991; http://alces.med.umn.edu/candida/codons.html;http://www.kazusa.or.jp/codon). To ensure a high expression of thesynthetic gene, the subset of ‘optimal’ codons of highly expressed geneswas used to design the synthetic BAX gene.

[0304] The synthCaBAX gene was constructed in three parts using eightoligonucleotides (FIG. 5). The sequences of the oligonucleotides aregiven in Table 7. Primer A1 introduced upstream of the ATG codon a Pst Isite and a Bgl II site. The Pst I site was used later on for directcloning into the Candida albicans expression vector, while the Bgl IIsite served as a linker for a yEGFP fusion. Primer C2 introduced a Sma Isite, suitable for cloning into the expression vector.

[0305] Fragment A and B were synthesised in two steps: in a first PCRround primer X1 and X2 (X represents A or B, respectively) were usedtogether. The resulting fragment served as a template in a second PCRround together with primers X1 and X3. Fragment C was synthesised in asingle PCR round using the primers C1 and C2. Fragment A and B werecloned into the pCR-BluntII-TOPO vector (Stratagene), while fragment Cwas cloned into the pCR2.1-TOPO vector (Stratagene). All three fragmentswere sequenced to ensure that no mutation was introduced by the PCR.

[0306] Subsequently, fragment A was digested with Pst I and Taq I,fragment B wit Taq I and Bam HI and fragment C with Bam HI and Sma I.The three products were cloned in a quadruple ligation into pUC21digested with Pst I and Sma I resulting in the plasmidpUC21:synthCandidaBAX. The sequence of the synthetic BAX gene is shownin FIG. 6.

[0307] Construction of Synthetic BAX- and GFP-Synthetic BAX ExpressionPlasmids

[0308] A Pst I-Sma I fragment containing the ORF of the synthetic BAXgene was cloned into the Pst I-Stu I digested vector pGAL1ACT1LUC (W.Martinet, EP application nr 99204557.5) resulting in the expressionconstruct pGAL1P:synthCaBAX (FIG. 7A). To facilitate recognition of theAUG codon during formation of initiation complexes a purine base (A) wasintroduced at position −3 from the AUG codon (Kozak 1981) using theQuick change site directed mutagenesis kit from Stratagene.

[0309] The yeast enhanced GFP gene yEGFP; (Cormack et al. 1997) wasamplified by PCR using primer 5′-AACTGCAGATGTCTAAAGGTGAAGMTTATTC-3′ (SEQID NO 11) as upstream primer and primer 5′-GGAAGATCTTCCTTTGTACAATTCATCCATACC-3′ (SEQ ID NO 12) as downstream primer. The sense primerintroduced a Pst I site (shown in bold and italic), while the anti-senseprimer contained a Bgl II linker (shown in bold and italic) for fusionwith the synthetic BAX gene. After cloning of the yEGFP gene into thepCR2.1-TOPO vector (Stratagene), the gene was sequenced to ensure thatno mutation was introduced by PCR.

[0310] The yEGFP-synth Candida BAX fusion was created by cloning aPstI-BglII yEGFP fragment together with a Bgl II-Sma I synthetic CandidaBAX fragment into the Pst I-Stu I digested expression vector pGAL1ACT1LUC. The obtained pGAL1 P:yEGFP-synthCaBAX fusion construct (FIG.7B) was sequenced to ensure that no frameshift had occurred.

[0311] Creation of the Synthetic BAX Expression Strains

[0312] Transformation of the expression plasmids was performed using amodified procedure (Logghe, unpublished) of the spheroblasting protocol(Herreros et al. 1992). The plasmids were linearised with Bpu1102 I toallow directed integration into the genome at the GALL promoter site.Correct integration was analysed by Southern blotting. Therefore genomicDNA from different transformants was prepared using the Nucleon®extraction and purification kit (Amersham Pharmacia Biotech) anddigested with Xba I. The BAX probe used in the Southern blot wasprepared by PCR. The PCR was performed using the pGAL1 P:synthCaBAXplasmid as template, together with the sense primer5′-ATGGATGGTTCTGGTGMC-3′ (SEQ ID NO 13) and the anti-sense primer5′-TTAACCCATTTTTTTCCAGATG-3′ (SEQ ID NO 14). Standard PCR conditionswere used. For detection of the yEGFP a probe was synthesised by PCRusing primer 5′-AGAGATCTCGAGGGATCC-3′ (SEQ ID NO 15) as sense primer andprimer 5′-GCATTATTTGTACMTTCATCC-3′ (SEQ ID NO 16) as anti-sense primer.Southern blot hybridisation and detection were performed using theAlkPhos DIRECT labelling and detection system (Amersham PharmaciaBiotech) following the instructions of the manufacturer.

[0313] Western Blot Analysis

[0314] For Western blot analysis cells were pre-grown over night inSD-ura media till late log phase. The cells were harvested bycentrifugation, washed twice with water and inoculated in SG-ura toinduce Bax expression. Induction was performed for 15 hours. Yeast crudeextracts were prepared as described before (Sambook, Fritsch et al.1989). Detection of the Bax protein was performed using a polyclonalrabbit anti-mouse/rat Bax antibody (Pharmingen). Due to contamination ofthis antibody with yeast cell wall mannan antibodies, a very highbackground occurred. This problem could be avoided by pre-incubation ofthe antibody with 0.5 mg/ml purified yeast mannan (Rossanese et al.1999). Detection of the Gfp protein was done using an anti-Gfpmonoclonal antibody (Molecular Probes, Eugene, Oreg., USA).

[0315] Growth Curves

[0316] For growth curves, yeast cells were grown for 24 h in SD-uramedium (supplemented with uridine for the wild type). These cultureswere harvested, washed twice with water and inoculated to an OD₆₀₀ of0.1 into fresh SD-ura or SG-ura media. Growth was monitored inmicrotitre plates using the Bioscreen C system (Labsystems).

[0317] Viability Tests

[0318] Cells were pregrown in minimal dextrose medium to an OD₆₀₀ of 1.After washing the cells twice with water they were switched to minimalmedium containing galactose as carbon source. At the time pointsindicated, samples were taken and equal cell amounts were spread onminimal dextrose plates. The appearing colonies represent the viablefraction of the pool.

[0319] Results: Conditional Expression of the Synthetic BAX Gene inCandida albicans

[0320] A cDNA encoding the full-length mouse Bax protein was placedunder control of the Candida albicans GALL promoter allowing forconditional expression when cells are grown in galactose containingmedia. Initial experiments were performed using the wild type mouse baxgene. Expression of this gene did not result in any detectablephenotype, no difference in growth compared to the wild type wasobserved when cells were grown on galactose containing media (data notshown). This could be due to the non-traditional codon strategy adoptedby Candida albicans and related species. Analysis of the codons used inthe mouse BAX gene revealed a for Candida albicans not optimal codonusage as found for highly expressed genes in this yeast. To ensure ahigh expression of the BAX gene a codon-adapted, synthetic version ofthe gene was created using the strategy described above. The syntheticBAX gene was fused to the yEGFP to allow screening for transformantswith a high yEGFP-synthCaBAX expression level using FACS technology. Thenewly obtained plasmids pGAL1 P:synthCaBAX and pGAL1:GFP-synthCaBAX weretransformed into the C. albicans CAI4 strain. Transformants wereselected on uridine-fee minimal medium. About 25 transformants of eachexpression construct were chosen and streaked onto minimal dextrosemedium (non-inducing conditions) as well as on minimal galactose medium(inducing conditions). After two days incubation at 30° C. alltransformants did grow on the glucose containing media. When galactosewas used as a sole carbon source, most of the transformants did not grow(FIG. 8). Southern blot analysis of the galactose negative transformantsrevealed that a copy of the synthCaBAX gene had been integrated into theendogenous copy of the GALL promoter. To study differences in growth,the transformants were grown over night in synthetic glucose containingmedium. Subsequently, cells were washed with water and switched to freshmedium containing galactose as carbon source. While the wild type straindid grow well on galactose containing media no growth was observed forthe Bax expressing transformants (FIGS. 9A and B). Western blot analysisof the synthCaBAX transformants showed accumulation of the Bax protein(15 hours Bax induction, FIG. 10). A similar result was observed whenimmunoblotting was performed with the GFP-synthCaBAX expressing strains.Here the fusion protein was detected at the expected molecular weight ofabout 45K under inducing conditions (galactose as carbon source). Inaddition to the fusion protein a band appeared at the molecular weightof about 20K. This corresponds to the molecular weight of the Gfpprotein alone. Addition of a Gfp-expressing strain as a positive controlto the western blot did confirm these results. Here the Gfp protein wasdetected at the same molecular weight as the unexpected band in theGFP-synthCaBAX expressing strain (FIG. 11). This is most probably due toa partly proteolytic degradation of the fusion protein. Analysis of theGfp-fluorescence using FACS technology showed a high Gfp-fluorescencesignal for the transformants expressing the fusion protein (FIG. 12).When cell viability was analysed, different results were obtained forthe synthCaBAX strain and the GFP-synthCaBAX strain. The synthCaBAXstrain showed quite a rapid decrease in the amount of colony formingunits during the first 6 hours of incubation on galactose containingmedia. Afterwards the process slowed down significantly. This is incontrast to the results obtained for the strain expressing thegfp-synthCabax fusion protein. Here almost all the cells died at a veryrapid rate during the first 3 hours of incubation in media containinggalactose as sole carbon source. It is possible that the Bax trigger inthe synthCabax expressing cells is not strong enough to kill all cells.The cell has enough time to activate a sort of defence mechanism,possibly by proteolytic degradation of the Bax protein. The situation isdifferent for the fusion protein. Gfp is a very stable protein itself.Fusion of the Gfp to another protein could result in a stabilisation ofthis protein. It would be more resistant to proteolytic degradation.This would explain the situation for the Gfp-Bax fusion. The Gfp-Baxprotein is more protected from proteolytic degradation. Like that it isfor a longer period present in the cell. The death trigger is herewithstronger, so the cells die faster. The time that the cells have toactivate the proteolytic machinery is not sufficient for them tosurvive. TABLE 1 Oligo Sequence 5′ -> 3′ A1AACTGCAGGAAGATCTTCCATGGATGGTTCTGGTGAACAATTGGGTTCTGGTGG (SED ID NO 3)TCCAACCTCTTCTGAACAAATCATGAAAACCGGTGCTTTCTTGTTG A2TAGAAGCATCTTGTGGTGGTTGTTCCAAGGTCAATTCTGGGGTTTCACCAGCC (SEQ ID NO 4)ATTCTACCAGCTCTATCTTGGATGAAACCTTGCAACAAGAAAGCACC A3GGAATTCTCGACATCAGCGATCATTCTTTGCAATTCCATGTTAGAATCCAATTC (SEQ ID NO 5)ATCACCGATTCTTCTCAAACATTCAGACAATTTTTTGGTAGAAGCATCTTGTG B1GGAATTCGCTGATGTCGATACCGATTCTCCAAGAGAAGTCTTCTTCAGAGTCG (SED ID NO 6)CTGCTGATATGTTCGCTGATGGTAACTTCAACTG B2AATTCTGGGACTTTGGTACACAAAGCTTTCAAGACCAATTTAGAAGCGAAGTA (SEQ ID NO 7)GAACAAAGCGACGACTCTACCCCAGTTGAAGTTACCA B3CCACCTTGATCTTGGATCCAGACCAACAATCTTTCTCTCAAGAAATCCAAGGTC (SEQ ID NO 8)CAACCCATGATGGTTCTGATCAATTCTGGGACTTTG C1ATTGTTGGTCTGGATCCAAGATCAAGGTGGTTGGGAAGGTTTGTTGTCTTACTT (SEQ ID NO 9)CGGTACCCCAACCTGGCAAACCGTCA C2TCCCCCGGGGGATTAACCCATTTTTTTCCAGATGGTCAAAGAAGCGGTCAAGAC (SEQ ID NO 10)ACCAGCGACGAAGATGGTGACGGTTTGCCAGGTTGGG

[0321] TABLE 2 Overview of the differentially expressed genes after 30min Bax expression Comparison: INVSc1 YIpUTL versus INVSc1 YIpUTyLBNormalised intensities Up/ ORF Gene L YLB down Qt value Cellular role:Cell cycle control YBR133C HSL7 18932.54 37877.20 ↑ 2.00 Cellular role:Polymerase II transcription YDR253C MET32 17661.13 45567.17 ↑ 2.58YBR112C SSN6 26698.87 65315.83 ↑ 2.45 YDR145W TAF61 38697.96 73117.62 ↑1.89 YBR289W SNF5 33111.77 72328.70 ↑ 2.18 YDR216W ADR1 30127.45 8815.87↓ 3.42 YEL009C GCN4 16533.76 3030.44 ↓ 5.46 YBR089C-A NHP6B 22698.636297.49 ↓ 3.60 YMR043W MCM1 39141.64 84180.45 ↑ 2.15 YKR092C SRP405965.63 16105.82 ↑ 2.70 YMR273C ZDS1 14699.61 35508.04 ↑ 2.42 YPL089CRLM1 34922.91 67856.88 ↑ 1.94 YOR372C NDD1 20285.12 44445.20 ↑ 2.19YPL037C EGD1 30633.33 5250.70 ↓ 5.83 Cellular role: Cell polarityYBL085W BOI1 7693.29 18614.99 ↑ 2.42 Cellular role: Chromatine structureYBR009C HHF1 16668.00 4178.80 ↓ 3.99 YNL030W HHF2 49878.04 12566.96 ↓3.97 YDR224C HTB1 67355.40 23156.82 ↓ 2.91 YBL002W HTB2 25269.02 5383.97↓ 4.69 Cellular role: RNA processing YER112W USS1 12776.74 31470.70 ↑2.46 YPL190C NAB3 6381.36 17892.11 ↑ 2.80 YNL112W DBP2 9956.84 28036.48↑ 2.82 Cellular role: Energy generation YPL078C ATP4 26902.69 5980.38 ↓4.50 YDL004W ATP16 36525.08 3004.34 ↓ 12.16 YDR377W ATP17 14419.41756.86 ↓ 19.05 YDR529C QCR7 35346.95 5394.65 ↓ 6.55 YGR008C STF213275.51 2276.27 ↓ 5.83 YEL039C CYC7 13604.38 2689.66 ↓ 5.06 YKL150WMCR1 105337.67 30743.75 ↓ 3.43 YLR038C COX12 52687.73 5455.83 ↓ 9.66YLR327C 113.966.77 54.014.65 ↓ 2.11 Cellular role: Carbohydratemetabolism YBR149W ARA1 15149.55 4095.17 ↓ 3.70 YHR094C HXT1 12526.90785.73 ↓ 15.94 YDR345C HXT3 36643.13 1632.48 ↓ 22.45 YDR343C HXT677064.71 32060.05 ↓ 2.40 YDR342C HXT7 76349.13 27615.15 ↓ 2.76 Cellularrole: Signal transduction YER177W BMH1 22856.29 44771.71 ↑ 1.96 YDR099WBMH2 40127.38 74572.38 ↑ 1.86 YGR070W ROM1 12055.28 28169.57 ↑ 2.34YGR023W MTL1 7354.78 19648.06 ↑ 2.67 Cellular role: Protein synthesisYGR034W RPL26B 71942.48 74625.22 ↑ 1.04 Cellular role: Protein foldingYLR216C CPR6 9616.80 31126.02 ↑ 3.24 Cellular role: Proteinmodification/degradation YFR052W RPN12 5583.57 14855.67 ↑ 2.66 YDL147WRPN5 31932.20 52939.11 ↑ 1.66 YGR132C PHB1 15429.56 5591.19 ↓ 2.76YGR135W PRE9 39921.63 5517.17 ↓ 7.24 YFR010W UBP6 1892.76 828.94 ↓ 2.28Cellular role: Cell stress YIR037W GPX3 7869.22 21789.00 ↑ 2.77 YDR513WTTR1 55986.32 33263.12 ↓ 1.68 YCL035C GRX1 70248.30 10969.97 ↓ 6.40YFL014W HSP12 41689.29 18658.48 ↓ 2.23 YHR053C CUP1A 72852.07 43488.52 ↓1.68 YHR055C CUP1B 71934.03 56799.80 ↓ 2.77 YMR173W DDR48 16670.705022.40 ↓ 3.32 YMR251W- HOR7 26879.95 417.36 ↓ 64.41 A YLR043C TRX158251.39 4435.79 ↓ 13.13 YBL064C PRX1 21525.00 40969.00 ↑ 1.90 YOL151WGRE2 2624.55 24152.03 ↑ 9.20 Cellular role: Unknown YBL081W 73834.1174612.35 ↑ 1.01 YDR366C 39998.46 57428.80 ↑ 1.44 YCR004C YCP4 6869.0628115.73 ↑ 4.09 YCR013C 3988.55 15144.34 ↑ 3.80 YBR050C REG2 4687.9114408.20 ↑ 3.07 YBL109W 18744.60 35440.24 ↑ 1.89 YDR154C 19565.2369428.03 ↑ 3.55 YEL071W DLD3 22235.73 68790.83 ↑ 3.09 YHR095W 14426.7634896.68 ↑ 2.42 YGR069W 43413.57 72420.39 ↑ 1.67 YDR544C 13567.0027004.37 ↑ 1.99 YGR236C 24927.59 8032.35 ↓ 3.10 YIL057C 24246.39 773.56↓ 31.34 YGL080W 23425.00 3217.81 ↓ 7.28 YGL072C 16437.52 2652.80 ↓ 6.20YHR056C RSC30 72072.88 57446.85 ↓ 1.25 YKL054C VID31 17990.49 38258.80 ↑2.13 YLR311C 7992.40 24164.87 ↑ 3.02 YJR115W 64690.69 102066.34 ↑ 1.58YJL188C BUD19 7580.28 22325.70 ↑ 2.95 YKR040C 50934.78 100733.41 ↑ 1.98YLR053C 8117.66 20317.34 ↑ 2.50 YOR121C 59950.94 92470.43 ↑ 1.54 YNL143C98911.28 110534.34 ↑ 1.12 YOR131C 7941.55 22353.72 ↑ 2.81 YNL338W21800.45 38777.28 ↑ 1.78 YNL179C 13729.36 39516.53 ↑ 2.88 YOL150C3408.74 60298.39 ↑ 17.69 YMR107W 65118.70 10042.46 ↓ 6.48 YKL065C YET169556.19 12804.88 ↓ 5.43 YJR096W 21780.37 10655.13 ↓ 2.04 YJL161W16468.73 2618.26 ↓ 6.29 YML128C MSC1 80130.20 13795.84 ↓ 5.81 YMR251W26879.95 417.36 ↓ 64.41 YMR173W- 110104.98 61951.23 ↓ 1.78 A YPL201C17913.32 5018.97 ↓ 3.57 YOR285W 64074.73 29749.43 ↓ 2.15 YOR286W13458.08 733.06 ↓ 18.36 Cellular role: Cell wall maintenance YKR076WECM4 2674.15 13040.04 ↑ 4.88 YLR390W ECM19 5472.05 15145.85 ↑ 2.77Cellular role: Membrane fusion YHR138C 19921.35 3707.57 ↓ 5.37 Cellularrole: Vesicular transport YHR161C YAP180A 13086.35 30160.90 ↑ 2.30YPL085W SEC16 6668.57 15206.49 ↑ 2.28 YKL196C YKT6 18933.84 2890.07 ↓6.55 YPR028W YIP2 25434.34 2049.47 ↓ 12.41 Cellular role: DNArepair/recombination YDL059C RAD59 1948.61 13089.13 ↑ 6.72 Cellularrole: DNA synthesis YEL032W MCM3 23422.85 44327.48 ↑ 1.89 Cellular role:Amino acid metabolism YIL074C SER33 3978.42 16702.66 ↑ 4.20 YGR155W CYS44184.59 19270.89 ↑ 4.61 Cellular role: Fatty acid metabolism YHR179WOYE2 2291.36 40274.02 ↑ 17.58 Cellular role: Protein translocationYNL131W TOM22 16287.21 1679.78 ↓ 9.70 Cellular role: Small moleculetransport YDR276C SNA1 21148.46 1580.68 ↓ 13.38 YOR267C HRK1 62689.30110516.24 ↑ 1.76 YHR039-C VMA10 60107.90 8490.93 ↓ 7.08 YOR382W FIT26780.82 27236.15 ↑ 4.02

[0322] TABLE 3 Overview of the differentially expressed genes after 1 hBax expression Comparison: INVSc1 YIpUTL versus INVSc1 YIpUTyLBNormalised intensities Up/ ORF Gene L YLB down Qt value Cellular role:Polymerase II transcription YDR145W TAF61 20729.58 57376.27 ↑ 2.77YDR216W ADR1 5925.91 18459.00 ↑ 3.11 YBR112C CYC8 50186.77 64511.50 ↑1.29 YMR043W MCM1 21011.54 53700.49 ↑ 2.56 YPL089C RLM1 23440.5464284.32 ↑ 2.74 YOR372C NDD1 26412.58 50804.99 ↑ 1.92 Cellular role:Cell cycle control YBR133C HSL7 18761.64 53238.86 ↑ 2.84 Cellular role:Cell polarity YBL085W BOI1 37895.40 57761.52 ↑ 1.52 Cellular role:Chromatine structure YDR224C HTB1 13661.40 55656.34 ↑ 4.07 Cellularrole: Energy generation YGR183C QCR9 23181.54 81865.40 ↑ 3.53 YLR294C5054.57 28994.72 ↑ 5.74 YKL150W MCR1 43663.07 60593.16 ↑ 1.39 YMR256CCOX7 7606.58 28801.54 ↑ 3.79 YOL126C MDH2 34144.61 65326.97 ↑ 1.91YLR327C 97415.94 101651.17 ↑ 1.04 Cellular role: Vesicular transportYHR161C YAP180A 11602.81 34695.20 ↑ 2.99 YLR206W ENT2 14439.24 34621.70↑ 2.40 Cellular role: Carbohydrate metabolism YDR342C HXT7 65273.5622231.06 ↓ 2.94 YDR343C HXT6 43572.28 6075.38 ↓ 7.17 YDR345C HXT376352.52 40296.00 ↓ 1.89 YGR192C TDH3 38472.30 14145.84 ↓ 2.72 YKR097WPCK1 22919.81 38225.98 ↑ 1.67 YOR374W ALD4 33711.37 2607.43 ↓ 12.93Cellular role: Signal transduction YER177W BMH1 16298.14 31748.91 ↑ 1.95YDR099W BMH2 50572.45 65123.58 ↑ 1.29 Cellular role: Cell wallmaintenance YLR110C CCW12 102525.29 11230.41 ↓ 9.13 Cellular role:Protein modification/degradation YOR261C RPN8 12575.49 32568.47 ↑ 2.59Cellular role: Cell stress YHR053C CUP1A 32531.53 63579.94 ↑ 1.95YHR055C CUP1B 27939.92 65142.82 ↑ 2.33 YMR173W DDR48 38338.83 60514.70 ↑1.58 YOR031W CRS5 2922.32 23848.60 ↑ 8.16 YLR109W AHP1 43067.08 6302.46↓ 6.83 Cellular role: Unknown YBL081W 82476.13 44279.86 ↑ 1.86 YBL109W22998.63 63428.23 ↑ 2.76 YDR366C 14599.17 46494.73 ↑ 3.18 YDR154C21296.57 56534.93 ↑ 2.65 YGR236C SPG1 17717.80 64439.96 ↑ 3.64 YHR056CRSC30 27020.16 65110.42 ↑ 2.41 YGR182C 8171.02 34669.96 ↑ 4.24 YDR544C14797.70 37704.91 ↑ 2.55 YHR162W 13836.79 33381.64 ↑ 2.41 YGR243W30829.66 59765.39 ↑ 1.94 YBR050C REG2 14008.24 29603.16 ↑ 2.11 YEL071WDLD3 19487.41 35273.39 ↑ 1.81 YDR133C 83074.54 62986.96 ↓ 1.32 YDR134C83111.03 16839.53 ↓ 4.94 YHL021C 46028.06 8577.00 ↓ 5.37 YKL054C VID3128018.46 66537.91 ↑ 2.37 YLR311C 7803.52 31160.73 ↑ 3.99 YMR107W13453.15 78850.98 ↑ 5.86 YKL066W 8751.84 24129.32 ↑ 2.76 YMR173W-A38338.83 60514.70 ↑ 1.58 YML053C 23670.86 66254.48 ↑ 2.80 YOR121C17039.58 58016.58 ↑ 3.40 YOL106W 19917.67 69853.66 ↑ 3.51 YNL338W17864.90 49911.08 ↑ 2.79 YJR115W 84858.02 98161.71 ↑ 1.16 Cellular role:Small molecule transport YOR267C HRK1 90123.84 96824.51 ↑ 1.07

[0323] TABLE 4 Overview of the differentially expressed genes after 2 hBax expression Comparison: INVSc1 YIpUTL versus INVSc1 YIpUTyLBNormalised intensities Up/ Qt ORF Gene L YLB Down value Cellular role:Protein modification/degradation YCL052C PBN1 5261.22 8175.70 ↑ 1.55YDL147W RPN5 22386.40 47857.67 ↑ 2.14 YOR261C RPN8 27349.25 42198.05 ↑1.54 YGR132C PHB1 5252.03 8459.53 ↑ 1.61 YBR139W 9458.26 3611.21 ↓ 2.62Cellular role: Unknown YDR202C RAV2 7483.71 10089.19 ↑ 1.35 YBR062C4893.97 9894.82 ↑ 2.02 YDR366C 25468.2 59682.92 ↑ 2.34 YBL109W 24803.6237444.64 ↑ 1.51 YDR154C 21166.26 33434.35 ↑ 1.58 YEL071W DLD3 34153.8544083.39 ↑ 1.29 YGR236C SPG1 16978.52 31419.12 ↑ 1.85 YGR182C 30569.3158805.05 ↑ 1.92 YDR544C 15937.14 24421.99 ↑ 1.53 YHR162W 26610.3433794.73 ↑ 1.27 YHR056C RSC30 33372.66 68425.24 ↑ 2.05 YDR133C 75520.9962984.59 ↓ 1.20 YCR010C ADY2 17240.59 11835.82 ↓ 1.46 YDR134C 72723.669776.23 ↓ 7.44 YGR069W 65418.73 53767.35 ↓ 1.22 YIL057C 16510.16 2198.04↓ 7.51 YGL072C 12209.68 6509.91 ↓ 1.88 YGL080W 22550.76 11525.24 ↓ 1.96YLR311C 11095.31 24660.47 ↑ 2.22 YJR115W 74757.79 103422.48 ↑ 1.38YMR099C 7057.15 11477.42 ↑ 1.63 YMR173W-A 31901.05 48886.91 ↑ 1.47YML132W COS3 24648.97 34895.33 ↑ 1.42 YKL066W 13581.94 25433.97 ↑ 1.87YJL142C 7205.86 11920.21 ↑ 1.65 YLR346C 6447.57 11569.63 ↑ 1.79 YLR053C41161.10 78636.82 ↑ 1.91 YMR110C 19410.64 29661.23 ↑ 1.53 YKR075C19104.57 29948.72 ↑ 1.57 YOR121C 36492.56 59452.09 ↑ 1.63 Cellular role:Unknown YOL106W 31382.10 76664.72 ↑ 2.44 YNL338W 24117.93 38981.22 ↑1.62 YNL134C 9617.33 14613.60 ↑ 1.52 YKL065C YET1 52422.65 33794.03 ↓1.55 YMR009W 20666.22 9519.29 ↓ 2.17 YJL144W 10316.92 3122.77 ↓ 3.30YML128C MSC1 584128.13 25434.11 ↓ 2.29 YNL179C 21938.96 10883.98 ↓ 2.02YOL109W ZEO1 22711.98 6581.11 ↓ 3.45 YNR002C FUN34 18241.25 9752.25 ↓1.87 Cellular role: Chromatine structure YDR224C HTB1 25356.73 30827.54↑ 1.22 YBL002W HTB2 9241.68 14261.54 ↑ 1.54 YBL003C HTA2 3453.55 6553.49↑ 1.90 YNL031C HHT2 13376.02 2348.84 ↓ 5.69 Cellular role: Polymerase IItranscription YBR289W SNF5 59542.27 65885.13 ↑ 1.11 YDR073W SNF1112190.01 23088.03 ↑ 1.89 YMR043W MCM1 66457.16 77022.05 ↑ 1.16 YPL089CRLM1 49844.99 60624.28 ↑ 1.22 Cellular role: Signal transduction YDR099WBMH2 55902.13 73874.51 ↑ 1.32 Cellular role: Cell stress YBL064C PRX111203.87 14815.42 ↑ 1.32 YBR101C 25016.27 35781.64 ↑ 1.43 YLR043C TRX110864.53 3912.03 ↓ 2.78 YGR209C TRX2 30492.33 37829.20 ↑ 1.24 YER103WSSA4 8763.38 15799.18 ↑ 1.80 YHR055C CUP1B 18824.43 77613.05 ↑ 4.12YHR053C CUP1A 32726.62 63536.72 ↑ 1.94 YDR256C CTA1 9614.29 4232.17 ↓2.27 YCR021C HSP30 8090.05 3604.78 ↓ 2.24 YCL035C GRX1 28437.57 12843.99↓ 2.21 YGR086C 36796.12 24272.57 ↓ 1.52 YFL014W HSP12 61868.64 23288.19↓ 2.66 YOR031W CRS5 6015.69 14519.12 ↑ 2.41 YMR251W-A HOR7 17731.144231.39 ↓ 4.19 YOR120W GCY1 114252.98 78052.05 ↓ 1.46 Cellular role:Protein synthesis YAL003W EFB1 3044.80 5772.68 ↑ 1.90 YOL127W RPL256266.96 12055.41 ↑ 1.92 YHR010W RPL27 4057.16 10856.34 ↑ 2.68 YLR325CRPL38 5401.85 12955.89 ↑ 2.40 YJL189W RPL39 2044.64 8010.67 ↑ 3.92YIL148W RPL40A 5052.35 11595.54 ↑ 2.30 YKR094C RPL40B 3994.57 10011.13 ↑2.54 YOL139C CDC33 4132.18 8956.14 ↑ 2.17 Cellular role: Protein foldingYLR216C CPR6 20353.43 32713.37 ↑ 1.61 YKL117W SBA1 11144.25 1500.56 ↓7.43 Cellular role: Vesicular transport YCR009C RVS161 5350.32 9780.92 ↑1.83 YHR161C YAP180A 25136.63 32461.67 ↑ 1.29 YBL078C AUT7 16528.919843.25 ↓ 1.68 Cellular role: Carbohydrate metabolism YBL058W SHP14626.50 8179.94 ↑ 1.77 YBR149W ARA1 30706.41 9637.76 ↓ 3.19 YDR178W SDH414880.91 6237.35 ↓ 2.39 YHR094C HXT1 30389.99 18383.00 ↓ 1.65 YMR011WHXT2 39524.90 21221.96 ↓ 1.86 YDR345C HXT3 77025.40 56749.40 ↓ 1.36YDR343C HXT6 73149.70 8676.17 ↓ 8.43 YDR342C HXT7 75331.76 27052.43 ↓2.78 YKL060C FBA1 16273.54 21323.23 ↑ 1.31 Cellular role: Cell cyclecontrol YBR133C HSL7 32903 41964.32 ↑ 1.28 Cellular role: Energygeneration YMR256C COX7 18558.01 40422.91 ↑ 2.18 YML129C COX14 11418.5421798.88 ↑ 1.91 YFR033C QCR6 9159.48 13398.67 ↑ 1.46 YDR529C QCR724821.75 16556.87 ↓ 1.50 YJL166W QCR8 15554.30 24509.26 ↑ 1.58 YHR001W-AQCR10 12416.35 23465.31 ↑ 1.89 YBR039W ATP3 11709.79 3088.19 ↓ 3.79YPL078C ATP4 11325.64 13769.72 ↑ 1.22 YPL271W ATP15 3261.75 7839.05 ↑2.40 YLR327C 51742.90 128511.27 ↑ 2.48 YLR294C 15832.61 38544.44 ↑ 2.43YAL060W FUN49 11792.72 5778.91 ↓ 2.04 Cellular role: Small moleculetransport YDR276C SNA1 19337.39 12392.29 ↓ 1.56 YGR197C SNG1 4766.1810484.09 ↑ 2.20 YHR039C-B VMA10 21190.93 10592.98 ↓ 2.00 YOR267C HRK1111849.17 101339.10 ↓ 1.10 Cellular role: RNA processing YGR250C 8709.9217358.43 ↑ 1.99 Cellular role: Cell wall maintenance YER150W SP1155592.73 22403.59 ↓ 2.48 YLR110C CCW12 35147.41 5786.88 ↓ 6.07 Cellularrole: Cell polarity YOR122C PFY1 14459.45 20176.41 ↑ 1.40 Cellular role:Amino acid metabolism YPR035W GLN1 20894.14 7522.05 ↓ 2.78

[0324] TABLE 5 Overview of the differentially expressed genes after 3 hBax expression Comparison: INVSc1 YIpUTL versus INVSc1 YIpUTyLBNormalised intensities Up/ ORF Gene L YLB down Qt value Cellular role:Cell cycle control YBR133C HSL7 63562.10 43191.28 ↓ 1.47 Cellular role:Cell polarity YBL085W BOI1 32734.79 23497.41 ↓ 1.39 Cellular role:Chromatine structure YDR545W YRF1-1 20111.51 11479.67 ↓ 1.75 Cellularrole: Energy generation YCR005C CIT2 11882.42 25632.94 ↑ 2.16 YGR183CQCR9 74474.20 11510.99 ↓ 6.47 YOL126C MDH2 55984.88 17978.10 ↓ 3.11Cellular role: Carbohydrate metabolism YBR019C GAL10 3092.50 15697.54 ↑5.08 YDR345C HXT3 14086.41 25657.66 ↑ 1.82 YKR097W PCK1 50736.4420858.02 ↓ 2.43 Cellular role: Signal transduction YDR099W BMH2 63285.1656028.91 ↓ 1.13 Cellular role: Protein synthesis YHR010W RPL27A 23254.907217.14 ↓ 3.22 YLR325C RPL38 26725.96 9121.29 ↓ 2.93 Cellular role: Cellstress YFL014W HSP12 40848.44 69781.91 ↑ 1.71 YHR053C CUP1A 20399.1065037.14 ↑ 3.19 YHR055C CUP1B 21763.09 64594.58 ↑ 2.97 YMR173W DDR4875407.16 36354.37 ↓ 2.07 YOL052C-A DDR2 20479.72 33702.23 ↑ 1.65Cellular role: Unknown YIL057C 7602.78 24104.02 ↑ 3.17 YHR056C RSC3041473.41 64809.08 ↑ 1.56 YDR544C 55075.67 29731.72 ↓ 1.85 YKR040C48049.71 59649.47 ↑ 1.24 YNL338W 86107.91 30045.62 ↓ 2.87 YJR115W74889.58 81238.98 ↓ 1.08 YBL109W 64754.79 57185.99 ↓ 1.13 YMR173W-A75407.16 36354?37 ↓ 2.07

[0325] TABLE 6 Overview of the differentially expressed genes after 6 hBax expression Comparison: INVSc1 YIpUTL versus INVSc1 YIpUTyLBNormalised Intensities Up/ ORF Gene L YLB down Qt value Cellular role:Cell stress YDR171W HSP42 13484.04 27183.07 ↑ 2.02 YFL014W HSP1241197.12 29081.08 ↓ 1.42 YDR513W TTR1 19985.22 12935.62 ↓ 1.54 YCL035CGRX1 31735.39 12930.71 ↓ 2.45 YGR209C TRX2 54455.65 47569.21 ↓ 1.14YHR053C CUP1A 81488.84 15289.39 ↓ 5.33 YHR055C CUP1B 81278.95 20031.69 ↓4.06 YMR251W-A HOR7 18824.54 5914.28 ↓ 3.18 Cellular role: Signaltransduction YDR099W BMH2 29412.99 58598.42 ↑ 1.99 Cellular role:Protein synthesis YGL147C RPL9A 13655.66 1585.97 ↓ 8.61 YGR085C RPL11B27465.15 3791.35 ↓ 7.24 YDR418W RPL12B 14417.77 1555.24 ↓ 9.27 YLR029CRPL15A 37122.11 9321.81 ↓ 3.98 YOR312C RPL20B 50334.94 5706.59 ↓ 8.82YBR191W RPL21A 21740.90 2571.30 ↓ 8.46 YPL079W RPL21B 31059.43 5023.61 ↓6.18 YOL127W RPL25 75971.72 11749.17 ↓ 6.47 YHR010W RPL27A 45716.648096.40 ↓ 5.65 YDR471W RPL27B 14636.79 2613.40 ↓ 5.60 YDL075W RPL31A11969.47 2611.53 ↓ 4.58 YBL092W RPL32 7872.80 857.85 ↓ 9.18 YDL191WRPL35A 28582.59 6046.25 ↓ 4.73 YDL136W RPL35B 25433.49 5064.51 ↓ 5.02YLR325C RPL38 48051.23 8217.18 ↓ 5.85 YIL148W RPL40A 47028.95 9543.65 ↓4.93 YKR094C RPL40B 39900.50 5957.78 ↓ 6.70 YHR141C RPL42B 10163.88937.21 ↓ 10.84 YML063W RPS1B 15916.48 1144.54 ↓ 13.91 YGL123W RPS212505.56 2243.26 ↓ 5.57 YOR096W RPS7A 24164.37 3223.60 ↓ 7.50 YBL072CRPS8A 17198.50 3233.30 ↓ 5.32 YER102W RPS8B 16234.83 1791.18 ↓ 9.06YBR189W RPS9B 10075.22 2150.89 ↓ 4.68 YOR293W RPS10A 51787.23 12110.74 ↓4.28 YDR064W RPS13 9736.57 1587.67 ↓ 6.13 YDR450W RPS18A 37913.715674.60 ↓ 6.68 YML026C RPS18B 14458.01 2027.28 ↓ 7.13 YKL156W RPS27A23725.18 11117.26 ↓ 2.13 YLR167W RPS31 38648.54 2611.97 ↓ 14.80 YJL138CTIF2 20154.61 7264.66 ↓ 2.77 Cellular role: Energy metabolism YGR183CQCR9 57357.59 80447.53 ↑ 1.40 YDL004W ATP16 25047.95 10988.85 ↓ 2.28YKL150W MCR1 50931.46 37076.83 ↓ 1.37 YLR038C COX12 39506.06 29534.70 ↓1.34 Cellular role: Unknown YDR442W 14654.61 2242.42 ↓ 6.54 YDR134C17025.59 10561.72 ↓ 1.61 YHR056C RSC30 81350.52 31447.10 ↓ 2.59 YKR040C48390.21 90125.88 ↑ 1.86 YLR414C 13463.40 8085.92 ↓ 1.67 YLR312C25589.67 16184.57 ↓ 1.58 YJL188C BUD19 22074.09 4526.39 ↓ 4.88 YOR285W75099.98 61896.00 ↓ 1.21 YOL109W ZEO1 66287.15 35502.43 ↓ 1.87 Cellularrole: Chromatine structure YBR009C HHF1 11173.15 5416.74 ↓ 2.06 YNL030WHHF2 31366.74 20132.23 ↓ 1.56 Cellular role: Nucleotide metabolismYDR399W HPT1 13339.03 5333.81 ↓ 2.50 Cellular role: Polymerase IItranscription YEL009C GCN4 34617.98 20798.63 ↓ 1.66 YPL037C EGD117862.37 8229.01 ↓ 2.17 Cellular role: Vesicular transport YBL078C AUT742661.70 32333.01 ↓ 1.32 YOR327C SNC2 22716.56 13704.48 ↓ 1.66 Cellularrole: Small molecule transport YHR039C-B VMA10 44429.30 23826.51 ↓ 1.86Cellular role: Cell wall maintenance YKL097W-A CWP2 13529.93 1617.20 ↓8.37 Cellular role: Carbohydrate metabolism YKL060C FBA1 33329.7410367.82 ↓ 3.21

[0326] TABLE 7 Sequence ID NO ORF GENE 30 min 1 h 2 h 3 h 6 h SEQ ID NO17 YAL003W EFB1 1.90 SEQ ID NO 19 VAL060W FUN49 −2.00 SEQ ID NO 21YBL002W HTB2 −4.69 1.54 SEQ ID NO 23 YBL058W SHP1 1.77 SEQ ID NO 25YBL064C PRX1 1.90 1.32 SEQ ID NO 27 YBL072C RPS8A −5.32 SEQ ID NO 29YBL081W 1.01 1.86 SEQ ID NO 31 YBL085W BOI1 2.42 1.52 −1.39 SEQ ID NO 33YBL092W RPL32 2.76 −9.18 SEQ ID NO 35 YBL109W 1.89 2.76 1.51 −1.13 SEQID NO 37 YBR009C HHF1 −3.99 −2.06 SEQ ID NO 39 YBR019C GAL10 5.08 SEQ IDNO 41 YBR039W ATP3 −3.70 SEQ ID NO 43 YBR050C REG2 3.07 2.11 SEQ ID NO45 YBR062C 2.02 SEQ ID NO 47 YBR089C-A NHP6B −3.60 SEQ ID NO 49 YBR101C1.43 SEQ ID NO 51 YBR112C SSN6 2.45 1.29 SEQ ID NO 53 YBR133C HSL7 2.002.84 1.28 −1.47 SEQ ID NO 55 YBR139W −2.60 SEQ ID NO 57 YBR149W ARA1−3.70 −3.11 SEQ ID NO 59 YBR189W RPS9B −4.68 SEQ ID NO 61 YBR191W RPL21A−8.46 SEQ ID NO 63 YBR289W SNF5 2.18 1.11 SEQ ID NO 65 YCL035C GRX1−6.40 −2.20 −2.45 SEQ ID NO 67 YCL052C PBN1 1.55 SEQ ID NO 69 YCR004CYCP4 4.09 SEQ ID NO 71 YCR005C CIT2 2.16 SEQ ID NO 73 YCR009C RVS1611.83 SEQ ID NO 75 YCR010C −1.40 SEQ ID NO 77 YCR013C 3.80 SEQ ID NO 79YCR021C HSP30 −2.20 SEQ ID NO 81 YDL004W ATP16 −12.16 −2.28 SEQ ID NO 83YDL059C RAD59 6.72 SEQ ID NO 85 YDL075W RPL31A −4.58 SEQ ID NO 87YDL147W RPN5 1.66 2.14 SEQ ID NO 89 YDR064W RPS13 −6.13 SEQ ID NO 91YDR073W SNF11 1.89 SEQ ID NO 93 YDR099W BMH2 1.86 1.29 1.32 −1.13 1.99SEQ ID NO 95 YDR133C −1.32 −1.20 SEQ ID NO 97 YDR134C −4.94 −7.40 −1.61SEQ ID NO 99 YDR145W TAF61 1.89 2.77 SEQ ID NO 101 YDR154C 3.55 2.651.58 SEQ ID NO 103 YDR171W HSP42 2.02 SEQ ID NO 105 YDR178W SDH4 −2.30SEQ ID NO 107 YDR202C RAV2 1.35 SEQ ID NO 109 YDR216W ADR1 −3.42 3.11SEQ ID NO 111 YDR224C HTB1 −2.91 4.07 1.22 SEQ ID NO 113 YDR253C MET322.58 SEQ ID NO 115 YDR256C CTA1 −2.20 SEQ ID NO 117 YDR276C SNA1 −13.38−1.50 SEQ ID NO 119 YDR342C HXT7 −2.76 −2.94 −2.70 SEQ ID NO 121 YDR343CHXT6 −2.40 −7.17 −8.40 SEQ ID NO 123 YDR345C HXT3 −22.45 −1.89 −1.301.82 SEQ ID NO 125 YDR366C 1.44 3.18 2.34 SEQ ID NO 127 YDR377W ATP17−19.05 SEQ ID NO 129 YDR399W HPT1 −2.50 SEQ ID NO 131 YDR418W RPL12B−9.27 SEQ ID NO 133 YDR513W TTR1 −1.68 −1.54 SEQ ID NO 135 YDR544C 1.992.55 1.53 −1.85 SEQ ID NO 137 YDR545W YRF1-1 −1.75 SEQ ID NO 139 YEL009CGCN4 −5.46 −1.66 SEQ ID NO 697 YEL032W MCM3 1.89 SEQ ID NO 141 YEL039CCYC7 −5.06 SEQ ID NO 143 YEL071W DLD3 3.09 1.81 1.29 SEQ ID NO 145YER103W SSA4 1.80 SEQ ID NO 147 YER112W USS1 2.46 SEQ ID NO 149 YER150WSPI1 −2.40 SEQ ID NO 151 YER177W BMH1 1.96 1.95 SEQ ID NO 153 YFR010WUBP6 −2.28 SEQ ID NO 155 YFR033C QCR6 1.46 SEQ ID NO 157 YFR052W RPN122.66 SEQ ID NO 159 YGL072C −6.20 −1.80 SEQ ID NO 161 YGL080W −7.28 −1.90SEQ ID NO 163 YGL123W RPS2 −5.57 SEQ ID NO 165 YGR008C STF2 −5.83 SEQ IDNO 167 YGR023W MTL1 2.67 SEQ ID NO 169 YGR034W RPL26B 1.04 SEQ ID NO 171YGR069W 1.67 −1.20 SEQ ID NO 173 YGR070W ROM1 2.34 SEQ ID NO 175 YGR086C−1.50 SEQ ID NO 177 YGR132C PHB1 −2.76 1.61 SEQ ID NO 179 YGR135W PRE9−7.24 SEQ ID NO 181 YGR155W CYS4 4.61 SEQ ID NO 183 YGR192C TDH3 −2.72SEQ ID NO 185 YGR197C SNG1 2.20 SEQ ID NO 187 YGR209C TRX2 1.24 −1.14SEQ ID NO 189 YGR243W 1.94 SEQ ID NO 191 YGR250C 1.99 SEQ ID NO 193YHL021C −5.37 SEQ ID NO 195 YHR001W-A QCR10 1.89 SEQ ID NO 197 YHR039C-BVMA10 −7.08 −2.00 −1.86 SEQ ID NO 199 YHR053C CUP1A −1.68 1.95 1.94 3.19−5.33 SEQ ID NO 201 YHR055C CUP1B −2.77 2.33 4.12 2.97 −4.06 SEQ ID NO203 YHR056C −1.25 2.41 2.05 1.56 −2.59 SEQ ID NO 205 YHR094C HXT1 −15.94−1.60 SEQ ID NO 207 YHR095W 2.42 SEQ ID NO 209 YHR138C −5.37 SEQ ID NO211 YHR161C YAP180A 2.30 2.99 1.29 SEQ ID NO 213 YHR162W 2.41 1.27 SEQID NO 215 YHR179W OYE2 17.58 SEQ ID NO 217 YIL057C −31.34 −7.50 3.17 SEQID NO 219 YIL074C SER33 4.20 SEQ ID NO 221 YIR037W GPX3 2.77 SEQ ID NO223 YJL138C TIF2 −2.77 SEQ ID NO 225 YJL142C 1.65 SEQ ID NO 227 YJL144W−3.30 SEQ ID NO 229 YJL161W −6.29 SEQ ID NO 231 YJL166W QCR8 1.58 SEQ IDNO 233 YJR096W −2.04 SEQ ID NO 235 YJR115W 1.58 1.16 1.38 −1.08 SEQ IDNO 237 YKL054C VID31 2.13 2.37 SEQ ID NO 239 YKL060C FBA1 1.31 −3.21 SEQID NO 241 YKL065C YET1 −5.43 −1.55 SEQ ID NO 243 YKL066W 2.76 1.87 SEQID NO 245 YKL097W-A CWP2 −8.37 SEQ ID NO 247 YKL117W SBA1 −7.43 SEQ IDNO 249 YKL150W MCR1 −3.43 1.39 −1.37 SEQ ID NO 251 YKL156W RPS27A −2.13SEQ ID NO 253 YKL196C YKT6 −6.55 SEQ ID NO 255 YKR040C 1.98 1.24 1.86SEQ ID NO 257 YKR075C 1.57 SEQ ID NO 259 YKR076W ECM4 4.88 SEQ ID NO 261YKR092C SRP40 2.70 SEQ ID NO 263 YKR097W PCK1 1.67 −2.43 SEQ ID NO 265YLR029C RPL15A −3.98 SEQ ID NO 267 YLR038C COX12 −9.66 −1.34 SEQ ID NO269 YLR043C TRX1 −13.13 −2.78 SEQ ID NO 271 YLR053C 2.50 1.91 SEQ ID NO273 YLR109W AHP1 −6.83 SEQ ID NO 275 YLR110C −9.13 −6.07 SEQ ID NO 277YLR206W ENT2 2.40 SEQ ID NO 279 YLR216C CPR6 3.24 1.61 SEQ ID NO 281YLR294C 5.74 2.43 SEQ ID NO 283 YLR311C 3.02 3.99 2.22 SEQ ID NO 285YLR312C −1.58 SEQ ID NO 287 YLR327C −2.10 1.04 2.48 SEQ ID NO 289YLR346C 1.79 SEQ ID NO 291 YLR390W ECM19 2.77 SEQ ID NO 293 YLR414C−1.67 SEQ ID NO 295 YML053C 2.80 SEQ ID NO 297 YML129C COX14 1.91 SEQ IDNO 299 YML132W COS3 1.42 SEQ ID NO 301 YMR009W −2.17 SEQ ID NO 303YMR011W HXT2 −1.86 SEQ ID NO 305 YMR043W MCM1 2.15 2.56 1.16 SEQ ID NO307 YMR099C 1.63 SEQ ID NO 309 YMR107W −6.48 5.86 SEQ ID NO 311 YMR110C1.53 SEQ ID NO 313 YMR173W DDR48 −3.32 1.58 −2.07 SEQ ID NO 691YMR173W-A −1.78 1.58 1.47 −2.07 SEQ ID NO 315 YMR251W −64.41 SEQ ID NO317 YMR251W-A HOR7 −64.41 −4.19 −3.18 SEQ ID NO 319 YMR256C COX7 3.792.18 SEQ ID NO 321 YMR273C ZDS1 2.42 SEQ ID NO 323 YNL030W HHF2 −3.97−1.56 SEQ ID NO 325 YNL031C HHT2 −5.69 SEQ ID NO 327 YNL112W DBP2 2.82SEQ ID NO 329 YNL131W TOM22 −9.70 SEQ ID NO 331 YNL134C 1.52 SEQ ID NO333 YNL143C 1.12 SEQ ID NO 335 YNL179C 2.88 −2.02 SEQ ID NO 337 YNL338W1.78 2.79 1.62 −2.87 SEQ ID NO 339 YNR002C FUN34 −1.87 SEQ ID NO 709YOL052C-A DDR2 1.65 SEQ ID NO 341 YOL106W 3.51 2.44 SEQ ID NO 343YOL109W ZEO1 −3.45 −1.87 SEQ ID NO 345 YOL126C MDH2 1.91 −3.11 SEQ ID NO347 YOL139C CDC33 2.17 SEQ ID NO 349 YOL150C 17.69 SEQ ID NO 351 YOL151WGRE2 9.20 SEQ ID NO 353 YOR120W GCY1 −1.46 SEQ ID NO 355 YOR121C 1.543.40 1.63 SEQ ID NO 357 YOR122C PFY1 1.40 SEQ ID NO 359 YOR131C 2.81 SEQID NO 361 YOR261C RPN8 2.59 1.54 SEQ ID NO 363 YOR267C 1.76 1.07 −1.10SEQ ID NO 365 YOR285W −2.15 −1.21 SEQ ID NO 367 YOR286W −18.36 SEQ ID NO369 YOR327C SNC2 −1.66 SEQ ID NO 371 YOR372C NDD1 2.19 1.92 SEQ ID NO373 YOR374W ALD4 −12.93 SEQ ID NO 375 YOR382W 4.02 SEQ ID NO 377 YPL037CEGD1 −5.83 −2.17 SEQ ID NO 379 YPL078C ATP4 −4.50 1.22 SEQ ID NO 381YPL079W RPL21B −6.18 SEQ ID NO 383 YPL085W SEC16 2.28 SEQ ID NO 385YPL089C RLM1 1.94 2.74 1.22 SEQ ID NO 387 YPL190C NAB3 2.80 SEQ ID NO389 YPL201C −3.57 SEQ ID NO 391 YPL271W ATP15 2.40 SEQ ID NO 393 YPR028WYIP2 −12.41 SEQ ID NO 395 YPR035W GLN1 −2.78

[0327] TABLE 8 C. albicans 522 CDS's S. cerevisiae 11645 CDS's totalcodon chosen for codons used in total aa codons frequency: per thousandnumber synthCaBAX gene wt muBAX gene frequency: per thousand number AlaGCU 30.7 8686 x 6 21.1 118595 GCC 12.7 3582 4 12.6 70785 GCA 15.4 4357 216.2 91018 GCG 2 578 1 6.1 34546 Arg CGU 5.9 1682 1 6.5 36518 CGC 0.7204 1 2.6 14571 CGA 3.5 989 3 3 16957 CGG 0.8 220 3 1.7 9801 AGA 23.66673 x 1 21.3 119672 AGG 2.7 769 2 9.3 52057 Asn AAU 37.9 10731 x 1 36202351 AAC 18.7 5293 2 24.9 140194 Asp GAU 43.6 12323 x 5 37.8 212658GAC 14.7 4152 7 20.4 114451 Cys UGU 9.7 2757 x 1 8 44797 UGC 1.7 493 14.7 26357 Gln CAA 35.2 9964 x 1 27.5 154529 CAG 6.9 1948 8 12.2 68463Glu GAA 49.5 14001 X 3 45.9 257930 GAG 11.5 3252 10 19.1 107568 Gly GGU33.5 9492 x 2 23.9 134515 GGC 4.5 1281 7 9.7 54629 GGA 13.7 3874 2 10.961481 His GGG 7.7 2182 8 6 33627 CAU 14 3964 13.7 77260 CAC 5.8 1642 7.843878 Ile AUU 39.9 11281 3 30.2 169795 AUC 14.2 4005 x 7 17.1 96126 AUA12.3 3478 17.8 100027 Leu UUA 1 295 26.3 148133 UUG 36.1 10204 x 2 27.1152590 CUU 9.8 2777 2 12.2 68479 CUC 2.5 694 7 5.4 30218 CUA 4 1133 113.4 75414 Lys AAA 48.6 13760 x 2 42.1 236746 AAG 19.4 5477 6 30.8173174 Met AUG 18.4 5219 x 8 20.9 117410 Phe UUU 28.6 8100 4 26 146355UUC 15.9 4486 x 7 18.2 102389 Pro CCU 13.2 3722 1 13.6 76366 CCC 3.61027 5 6.8 38247 CCA 26.6 7531 x 18.2 102277 CCG 2.4 686 1 5.3 29758 SerCUG 3.1 875 9 10.4 58583 UCU 23.3 6595 x 1 23.6 132608 UCC 10.3 2928 414.2 79928 UCA 24.6 6955 18.8 105570 UCG 6.5 1836 1 8.6 48186 AGU 23.66673 14.2 79649 AGC 4.5 1269 5 9.7 54330 Thr ACU 30.7 8689 1 20.2 113634ACC 13.9 3928 x 8 12.6 70777 ACA 17.4 4928 5 17.7 99759 ACG 3.6 1019 1 844817 Trp UGG 11 3115 x 6 10.3 58092 Tyr UAU 24 6782 18.8 105489 UAC11.6 3280 x 2 14.7 82483 Val GUU 33.2 9391 1 22 123726 GUC 10.3 2927 x 311.6 65203 GUA 8 2265 11.8 66100 GUG 10 2842 7 10.7 60033

[0328] TABLE 9 Regulation of 23 selected “Bax.specific” functions ORFGene Control Bax H2O2 B vs C Cellular role: Amino-acid metabolismYOR302W YOR302W 11541.92 26806.35 8895.74 2.32Cellular role: Cell stress YML028W TSA1 12889.91 2166.45 11327.36 0.17Cellular role: Chromatin/chromosome structure YBR009C HHF1 2149.698655.43 2909.14 4.03 YDR224C HTB1 13661.40 55656.34 18829.27 4.07YNL030W HHF2 8676.99 19603.93 4732.39 2.26Cellular role: Energy generation YBL099W ATP1 2728.21 8786.71 1644.483.22 YGR183C QCR9 23181.54 81865.40 24053.00 3.53 YJL166W QCR8 5296.7118093.93 5001.65 3.42 YLR038C COX12 7336.65 19935.69 5118.43 2.72Cellular role: Signal transduction YHR135C YCK1 3939.64 8358.11 3707.172.12 YOL100W PKH2 2218.45 6088.96 2619.31 2.74 Cellular role:Transcription factor YDR216W ADR1 5925.91 18459.00 6434.43 3.11Cellular role: Unknown YDR504C YDR504C 2741.47 6908.49 2839.62 2.52YGR146C YGR146C 2099.74 5616.94 1303.89 2.68 YGR236C SPG1 17717.8064439.96 24134.29 3.64 YHR138C YHR138C 6218.30 14817.41 5220.50 2.38YJL142C YJL142C 6988.27 16006.02 6740.46 2.29 YKL123W YKL123W 2826.825952.34 2766.04 2.11 YLR414C YLR414C 4510.80 11867.69 3531.27 2.63YMR107W YMR107W 13453.15 78850.98 17417.00 5.86 YOL099C YOL099C 3690.4511604.72 5454.15 3.14 YPL201C YPL201C 15960.14 33633.74 7449.66 2.11YJL060W YJL060W 8798.50 2406.39 6356.11 0.27

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0 SEQUENCE LISTING The patent application contains a lengthy “SequenceListing” section. A copy of the “Sequence Listing” is available inelectronic form from the USPTO web site(http://seqdata.uspto.gov/sequence.html?DocID=20040161840). Anelectronic copy of the “Sequence Listing” will also be available fromthe USPTO upon request and payment of the fee set forth in 37 CFR1.19(b)(3).

1. An isolated nucleic acid representing a synthetic BAX-gene selectedfrom the group consisting of: a) a nucleic acid comprising a sequence asrepresented by SEQ ID NO 1, b) a nucleic acid comprising a fragment of asequence of SEQ ID NO 1 and encoding a functional fragment of thesequence represented by SEQ ID NO 2, c) a nucleic acid comprising asequence as represented in any of SEQ ID NOs 3 to 10, d) a nucleic acidwhich is more than 75% identical to the nucleic acid as represented bySEQ ID NO 1, or to a nucleic acid according to the nucleic acid asdefined in b) or c), and, e) a nucleic acid as defined in any one of (a)to (i) interrupted by intervening DNA sequences, or a nucleic acidrepresenting the complement of any of said nucleic acids as defined in(a) to (d).
 2. An isolated nucleic acid according to claim 1 which isDNA, cDNA, genomic DNA, synthetic DNA, or RNA wherein T is replaced byU.
 3. A vector comprising a nucleic acid as defined in claim 1 or
 2. 4.A vector according to claim 3 which is an expression vector wherein saidnucleic acid sequence is operably linked to one or more controlsequences allowing the expression in prokaryotic and/or eukaryotic hostcells.
 5. An expression vector according to claim 4 which comprises aninducible promoter
 6. An expression vector according to claim 4 or 5which comprises a sequence encoding a reporter molecule.
 7. A vectoraccording to any of claims 3 to 6 for inducing programmed cell death inCandida spp.
 8. A host cell transformed, transfected or infected with avector according to any of claims 3 to
 7. 9. A host cell of claim 8which is a bacterial, yeast or fungal cell.
 10. A host cell according toclaim 8 or 9 wherein said cell is a Candida spp. cell.
 11. A geneticallymodified yeast or fungal cell according to claim 9 wherein saidmodification results in the onset of at least one pathway eventuallyleading to programmed cell death.
 12. A genetically modified Candidaspp. cell according to claim 10 wherein said modification results in theonset of at least one pathway eventually leading to programmed celldeath.
 13. A method for identifying Bax-resistant yeast or fungicomprising the steps of: a) providing (a) genetically modified yeast orfungi according to claim 11, b) treating said genetically modified yeastor fungi with a mutagen, c) isolating resistant yeast or fungal cells,and, d) optionally identifying and/or characterizing mutated genes Insaid resistant yeast or fungal cells.
 14. A method for identifyingCandida spp. sequences which are differentially expressed in a pathwayeventually leading to programmed cell death using a nucleic acid asdefined in claim 1 or 2, a vector according to any of claims 3 to 7 or agenetically modified host cell according to claim
 10. 15. A method forobtaining and identifying Candida spp. sequences involved in a pathwayeventually leading to programmed cell death comprising the steps of: a)providing a two hybrid system wherein a polypeptide encoded by a nucleicacid according to claim 1 or a vector according to any of claims 3 to 7as a bait and a Candida spp. cDNA library as a prey are expressed, b)detecting an interaction between said polypeptide and a Candida spp.polypeptide encoded by said cDNA library, and, c) identifying saidCandida spp. polypeptide or cDNA.
 16. A method for identifyinginhibitors (or inhibitor sequences) of Bax-induced cell death comprisingthe steps of: a) providing a genetically modified organism according toclaim 10, b) expressing a cDNA library in said genetically modifiedorganism, and, c) identifying a polypeptide or a cDNA which expressionhas a beneficial effect on the survival and/or growth of saidgenetically modified organism.
 17. A method according to claim 16wherein said genetically modified organism is a Candida spp.
 18. Anisolated Candida spp. nucleic acid identifiable by any of the methods ofany of claims 12 to
 17. 19. An isolated Candida spp. nucleic acidaccording to claim 18 selected from: (a) a nucleic acid encoding aprotein having an amino acid sequence as represented in any of SEQ IDNOs 434, 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420,422, 424, 426, 428, 430, 432, 436, 438, 440, 442, 444, 446, 448, 450,452, 454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478,480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506,508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534,536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562,564, 566, 568, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580,582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608,610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636,638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664,666, 668, 670, 672, 674, 688, 718, 720, 722, 724, 726, 728, 730 and 732,or encoding a functional equivalent, derivative or bioprecursor of saidprotein, b) a nucleic acid encoding a protein having an amino acidsequence which is more than 70% similar to any of the amino acidsequences represented in SEQ ID NOs 434, 398, 400, 402, 404, 406, 408,410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 436, 438,440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466,468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494,496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522,524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550,552, 554, 556, 558, 560, 562, 564, 566, 568, 560, 562, 564, 566, 568,570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596,598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624,626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652,654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 688, 718, 720,722, 724, 726, 728, 730 and 732, c) a nucleic acid encoding a proteinhaving an amino acid sequence which is more than 70% identical to any ofthe amino acid sequences represented in SEQ ID NOs 434, 398, 400, 402,404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430,432, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460,462, 464, 466, 468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488,490, 492, 494, 496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516,518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544,546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 560, 562,564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590,592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618,620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646,648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674,688, 718, 720, 722, 724, 726, 728, 730 and 732, d) a nucleic acidcomprising a sequence as represented in any of SEQ ID NOs 433, 397, 399,401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427,429, 431, 435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457,459, 461, 463, 465, 467, 469, 471, 473, 475, 477, 479, 481, 483, 485,487, 489, 491, 493, 495, 497, 499, 501, 503, 505, 507, 509, 511, 513,515, 517, 519, 521, 523, 525, 527, 529, 531, 533, 535, 537, 539, 541,543, 545, 547, 549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569,571, 573, 575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597,599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625,627, 629, 631, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653,655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 687, 717, 719, 721,723, 725, 727, 729 and 731, e) a nucleic acid which is more than 70%identical to any of the nucleic acid sequences as represented by any ofSEQ ID NOs 433, 397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417,419, 421, 423, 425, 427, 429, 431, 435, 437, 439, 441, 443, 445, 447,449, 451, 453, 455, 457, 459, 461, 463, 465, 467, 469, 471, 473, 475,477, 479, 481, 483, 485, 487, 489, 491, 493, 495, 497, 499, 501, 503,505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525, 527, 529, 531,533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557, 559,561, 563, 565, 567, 569, 571, 573, 575, 577, 579, 581, 583, 585, 587,589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615,617, 619, 621, 623, 625, 627, 629, 631, 633, 635, 637, 639, 641, 643,645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671,673, 687, 717, 719, 721, 723, 725, 727, 729 and 731, and f) a nucleicacid encoding a functional fragment of any of the nucleic acid sequencesas specified in any of a) to d),
 20. An isolated nucleic acid as definedin according to claim 19 which is DNA, cDNA, genomic DNA, synthetic DNA,or RNA wherein T is replaced by U.
 21. An isolated nucleic acid capableof selectively hybridizing to a nucleic acid as defined in any of claims18 to 20 or the complement thereof.
 22. An antisense molecule comprisinga nucleic acid capable of selectively hybridizing to a nucleic acid asdefined in any of claims 18 to
 21. 23. A nucleic acid probe whichselectively hybridises with any of the nucleic acid molecules as definedin claim 18 or
 19. 24. A nucleic acid primer which selectively amplifiesany of the nucleic acid molecules defined in claim 18 or
 19. 25. Anexpression vector comprising a nucleic acid according to any of claims18 to
 22. 26. An expression vector according to claim 25 which is anexpression vector wherein said nucleic acid is operably linked to one ormore control sequences allowing the expression in prokaryotic and/oreukaryotic host cells.
 27. An expression vector according to claim 25 or26 which comprises an inducible promoter.
 28. An expression vectoraccording to any of claims 25 to 27 which comprises a sequence encodinga reporter molecule.
 29. A host cell transformed, transfected orinfected with the vector of any of claims 25 to
 28. 30. An isolatednucleic acid according to any of claims 18 to 22 for use as amedicament.
 31. An isolated polypeptide which is involved in a pathwayfor programmed cell death of Candida spp. and encoded by a nucleic acidas defined in claim 18 or 19, wherein said polypeptide is selected from:(a) a polypeptide having an amino acid sequence as represented in any ofSEQ ID NOs 434, 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418,420, 422, 424, 426, 428, 430, 432, 436, 438, 440, 442, 444, 446, 448,450, 452, 454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476,478, 480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504,506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532,534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560,562, 564, 566, 568, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578,580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606,608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634,636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662,664, 666, 668, 670, 672, 674, 688, 718, 720, 722, 724, 726, 728, 730 and732, or encoding a functional equivalent, derivative or bioprecursor ofsaid protein; (b) a polypeptide having an amino acid sequence which ismore than 70% similar to any of the amino acid sequences as representedby any of SEQ ID NOs 434, 398, 400, 402, 404, 406, 408, 410, 412, 414,416, 418, 420, 422, 424, 426, 428, 430, 432, 436, 438, 440, 442, 444,446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468, 470, 472,474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500,502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528,530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556,558, 560, 562, 564, 566, 568, 560, 562, 564, 566, 568, 570, 572, 574,576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602,604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630,632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658,660, 662, 664, 666, 668, 670, 672, 674, 688, 718, 720, 722, 724, 726,728, 730 and 732, (c) a polypeptide having an amino acid sequence whichis more than 70% identical to any of the amino acid sequences asrepresented by any of SEQ ID NOs 434, 398, 400, 402, 404, 406, 408, 410,412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 436, 438, 440,442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468,470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496,498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524,526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552,554, 556, 558, 560, 562, 564, 566, 568, 560, 562, 564, 566, 568, 570,572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598,600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626,628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654,656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 688, 718, 720, 722,724, 726, 728, 730 and 732, and (d) a functional fragment of any of saidpolypeptides as defined in a) to c).
 32. A polypeptide according toclaim 31 for use as a medicament.
 33. An antibody capable ofspecifically binding to a polypeptide of claim 30 or to a specificepitope of said polypeptide.
 34. An antibody according to claim 33 foruse as a medicament.
 35. A pharmaceutical composition comprising anantibody of claim 33 or
 34. 36. Use of an isolated nucleic acid encodinga polypeptide which is involved in a pathway eventually leading toprogrammed cell death of yeast or fungi and which nucleic acid isselected from: (a) a nucleic acid encoding a protein having an aminoacid sequence as represented in any of SEQ ID NOs 18, 20, 22, 24, 26,28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62,64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98,100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126,128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154,156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182,184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210,212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238,240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266,268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294,296, 298, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312,314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 324, 326, 328, 340,342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368,370, 372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396,398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424,426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452,454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478, 480,482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506, 508,510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534, 536,538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564,566, 568, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582,584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610,612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638,640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666,668, 670, 672, 674, 688, 692, 694, 696, 698, 700, 702, 704, 706, 708,710, 712, 714, 716, 718, 720, 722, 724, 726, 728, 730 and 732, orencoding a functional equivalent, derivative or bioprecursor of saidprotein; (b) a nucleic acid encoding a protein having an amino acidsequence which is more than 70% similar to any of the amino acidsequences as represented by any of SEQ ID NOs 18, 20, 22, 24, 26, 28,30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64,66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100,102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128,130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156,158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184,186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212,214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240,242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268,270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296,298, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314,316, 318, 320, 322, 324, 326, 328, 330, 332, 324, 326, 328, 340, 342,344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370,372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396, 398,400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426,428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454,456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478, 480, 482,484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506, 508, 510,512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538,540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564, 566,568, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584,586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612,614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638, 640,642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668,670, 672, 674, 688, 692, 694, 696, 698, 700, 702, 704, 706, 708, 710,712, 714, 716, 718, 720, 722, 724, 726, 728, 730 and 732, (c) a nucleicacid encoding a protein having an amino acid sequence which is more than70% identical to any of the amino acid sequences as represented by anyof SEQ ID NOs 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44,46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80,82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112,114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140,142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168,170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196,198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224,226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252,254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280,282, 284, 286, 288, 290, 292, 294, 296, 298, 290, 292, 294, 296, 298,300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326,328, 330, 332, 324, 326, 328, 340, 342, 344, 346, 348, 350, 352, 354,356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382,384, 386, 388, 390, 392, 394, 396, 398, 400, 402, 404, 406, 408, 410,412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438,440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466,468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494,496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522,524, 526, 528, 530, 532, 534, 536, 538, 540 542, 544, 546, 548, 550,552, 554, 556, 558, 560, 562, 564, 566, 568, 560, 562, 564, 566, 568,570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596,598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624,626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652,654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 688, 692, 694,696, 698, 700, 702, 704, 706, 708, 710, 712, 714, 716, 718, 720, 722,724, 726, 728, 730 and 732, (d) a nucleic acid comprising a sequence asrepresented in any of SEQ ID NOs 17, 19, 21, 23, 25, 27, 29, 31, 33, 35,37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71,73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105,107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133,135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161,163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189,191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217,219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245,247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273,275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301,303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329,331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357,359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385,387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407, 409, 411, 413,415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441,443, 445, 447, 449, 451, 453, 455, 457, 459, 461, 463, 465, 467, 469,471, 473, 475, 477, 479, 481, 483, 485, 487, 489, 491, 493, 495, 497,499, 501, 503, 505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525,527, 529, 531, 533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553,555, 557, 559, 561, 563, 565, 567, 569, 571, 573, 575, 577, 579, 581,583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609,611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635, 637,639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665,667, 669, 671, 673, 687, 691, 693, 695, 697, 699, 701, 703, 705, 707,709, 711, 713, 715, 717, 719, 721, 723, 725, 727, 729 and 731, (e) anucleic acid which is more than 70% Identical to any of the nucleic acidsequences as represented by any of SEQ ID NOs 17, 19, 21, 23, 25, 27,29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63,65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99,101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127,129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155,157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183,185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211,213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239,241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267,269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295,297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323,325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351,353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379,381, 383, 385, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407,409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435,437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457, 459, 461, 463,465, 467, 469, 471, 473, 475, 477, 479, 481, 483, 485, 487, 489, 491,493, 495, 497, 499, 501, 503, 505, 507, 509, 511, 513, 515, 517, 519,521, 523, 525, 527, 529, 531, 533, 535, 537, 539, 541, 543, 545, 547,549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569, 571, 573, 575,577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603,605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631,633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659,661, 663, 665, 667, 669, 671, 673, 687, 691, 693, 695, 697, 699, 701,703, 705, 707, 709, 711, 713, 715, 717, 719, 721, 723, 725, 727, 729 and731, (f) a nucleic acid encoding a functional fragment of any of thenucleic acid sequences as specified in a) to e), and (g) the complementof any of the nucleic acid molecule as specified in a) to f), for thepreparation of a medicament for treating diseases associated with yeastor fungi.
 37. Use of an isolated polypeptide which is involved in apathway eventually leading to programmed cell death of yeast or fungi,said polypeptide being selected from: (a) a polypeptide having an aminoacid sequence as represented in any of SEQ ID NOs 18, 20, 22, 24, 26,28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62,64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98,100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126,128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154,156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182,184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210,212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238,240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266,268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294,296, 298, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312,314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 324, 326, 328, 340,342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368,370, 372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396,398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424,426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452,454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478, 480,482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506, 508,510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534, 536,538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564,566, 568, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582,584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610,612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638,640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666,668, 670, 672, 674, 688, 692, 694, 696, 698, 700, 702, 704, 706, 708,710, 712, 714, 716, 718, 720, 722, 724, 726, 728, 730 and 732, orencoding a functional equivalent, derivative or bioprecursor of saidprotein, (b) a polypeptide having an amino acid sequence which is morethan 70% similar o any of the amino acid sequences as represented by anyof SEQ ID NOs 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44,46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80,82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112,114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140,142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168,170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196,198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224,226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252,254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280,282, 284, 286, 288, 290, 292, 294, 296, 298, 290, 292, 294, 296, 298,300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326,328, 330, 332, 324, 326, 328, 340, 342, 344, 346, 348, 350, 352, 354,356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382,384, 386, 388, 390, 392, 394, 396, 398, 400, 402, 404, 406, 408, 410,412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438,440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466,468, 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494,496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522,524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550,552, 554, 556, 558, 560, 562, 564, 566, 568, 560, 562, 564, 566, 568,570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596,598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624,626, 628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652,654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 688, 692, 694,696, 698, 700, 702, 704, 706, 708, 710, 712, 714, 716, 718, 720, 722,724, 726, 728, 730 and 732, (c) a polypeptide having an amino acidsequence which is more than 70% identical to any of the amino acidsequences as represented by any of SEQ ID 18, 20, 22, 24, 26, 28, 30,32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66,68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100,102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128,130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156,158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 20, 178, 180, 182,184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210,212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238,240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266,268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294,296, 298, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312,314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 324, 326, 328, 340,342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368,370, 372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396,398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424,426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452,454, 456, 458, 460, 462, 464, 466, 468, 470, 472, 474, 476, 478, 480,482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502, 504, 506, 508,510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534, 536,538, 540 542, 544, 546, 548, 550, 552, 554, 556, 558, 560, 562, 564,566, 568, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582,584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610,612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, 636, 638,640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666,668, 670, 672, 674, 688, 692, 694, 696, 698, 700, 702, 704, 706, 708,710, 712, 714, 716, 718, 720, 722, 724, 726, 728, 730 and 732, and, (d)a functional fragment of any of said polypeptides as defined in a) toc), for the preparation of a medicament for treating diseases associatedwith yeast or fungi.
 38. A pharmaceutical or fungicidal compositioncomprising a nucleic acid as defined in claim 36 or a polypeptide asdefined in claim 37 together with a pharmaceutically acceptable carrierdiluent or excipient therefor.
 39. A vaccine for immunizing a mammalagainst yeast or fungal infections comprising at least one nucleic acidas defined in claim 36 or at least one polypeptide as defined in claim37 in a pharmaceutically acceptable carrier.
 40. A genetically modifiedyeast or fungus in which modification results in the overexpression orunderexpression of at least one of the nucleic acids as defined in claim36 or the polypeptides as defined in claim 36, which overexpression orunderexpression of said nucleic acid or polypeptide prevents, delays orsensitizes for apoptosis of said genetically modified yeast or fungus.41. A method of identifying compounds which selectively modulateexpression or functionality of polypeptides involved in a pathwayeventually leading to programmed cell death of yeast or fungi or inmetabolic pathways in which said polypeptides are involved, which methodcomprises: (a) contacting a compound to be tested with a geneticallymodified yeast or fungus according to claim 40, in addition tocontacting wild type cells with said compound, (b) monitoring the growthand/or death rate and/or activity of said genetically modified cellscompared to said wild type cells; wherein differential growth oractivity of said genetically modified yeast or fungi cells is indicativeof selective action of said compound on a polypeptide in the same or aparallel pathway, (c) alternatively monitoring the growth and/or deathrate and/or activity of said genetically modified cells compared togenetically modified cells which were not contacted with the compound tobe tested, wherein differential growth or activity of said geneticallymodified yeast of fungi cells is indicative of selective action of saidcompound on a polypeptide in the same or a parallel pathway, (d)alternatively monitoring changes in morphologic and/or functionalproperties of components in said genetically modified cells caused bythe addition of the compound to be tested, and, (e) identifying thecompound.
 42. A method of identifying compounds which selectivelymodulate expression or functionality of polypeptides involved in apathway eventually leading to programmed cell death of yeast and fungior in metabolic pathways in which said polypeptides are involved, whichmethod comprises: (a) contacting a compound to be tested with yeast orfungal cells transformed, transfected or infected with an expressionvector comprising an antisense sequence of at least one of the nucleicacid as defined in claim 36, which expression results in underexpressionof said polypeptide, in addition to contacting one or more wild typecells with said compound, (b) monitoring the growth and/or death rateand/or activity of said transformed, transfected or infected cellscompared to said wild type cells; wherein differential growth oractivity of said transformed, transfected or infected yeast or fungalcells is indicative of selective action of said compound on apolypeptide in the same or a parallel pathway, (c) alternativelymonitoring the growth and/or death rate and/or activity of saidtransformed, transfected or infected cells compared to transformed,transfected or infected cells which were not contacted with the compoundto be tested, wherein differential growth or activity of said mutatedyeast of fungi cells is indicative of selective action of said compoundon a polypeptide in the same or a parallel pathway, (d) alternativelymonitoring changes in morphologic and/or functional properties ofcomponents in said transformed, transfected or infected cells caused bythe addition of the compound to be tested, and, (e) identifying thecompound.
 43. A method of identifying compounds or polypeptides whichbind to or modulate the properties of polypeptides which are involved ina pathway eventually leading to programmed cell death of yeast or fungi,which method comprises: (a) contacting a compound or polypeptides to betested with at least one of the polypeptides as defined in claim 37, (b)detecting the complex formed between the compound or polypeptide to betested and said polypeptide, (c) alternatively, examining the diminutionof complex formation between said polypeptide and a binding partner,caused by the addition of the compound or polypeptide being tested, (d)alternatively, examining the alteration in the functional activity ofthe polypeptide, caused by the addition of the compound or polypeptidebeing tested, and, (e) identifying the compound or protein.
 44. A methodfor identifying compounds interacting with a polypeptide involved in apathway eventually leading to programmed cell death of yeast and fungicomprising the steps of: (a) providing a two-hybrid screening systemwherein a polypeptide of claim 37 and a protein interacting with saidpolypeptide or an interacting polypeptide obtainable by a method ofclaim 41, are expressed, (b) interacting said compound with the complexformed by the expressed proteins as defined in a), (c) detecting asecond complex, wherein the presence of said second complex identifies acompound which specifically binds to one of said polypeptide or to saidsecond complex, and, (d) identifying the compound.
 45. A method ofidentifying compounds which selectively modulate expression ofpolypeptides which are involved in a pathway eventually leading toprogrammed cell death of yeast or fungi which method comprises: (a)contacting host cells transformed, transfected or infected with anexpression vector comprising a promoter sequence of a nucleic acid asdefined in claim 36 joined in frame with a reporter gene, (b) monitoringincreased or decreased expression of said reporter gene caused by theaddition of the compound being tested, and, (c) identifying thecompound.
 46. A method for identifying polypeptides involved in apathway eventually leading to programmed cell death comprising the stepsof: (a) providing a two-hybrid system wherein a polypeptide encoded by anucleic acid according to claim 36 or a vector according to any ofclaims 3 to 7 as a bait and a yeast or fungal cDNA library as a prey areused, (b) detecting an interaction between said polypeptide and a yeastor fungal polypeptide encoded by said cDNA library, and, (c) identifyingsaid yeast or fungal polypeptide.
 47. A method according to any ofclaims 41 to 46 wherein said yeast or fungus is chosen fromSaccharomyces cerevisiae, Schizosaccharomyces pombe, Candida albicans,or Aspergillus fumigatus.
 48. A compound or polypeptide identifiableaccording to the method of any of claims 41 to
 47. 49. A compound orpolypeptide according to claim 48 for use as a medicament.
 50. A methodfor preparing a pharmaceutical composition for treating diseasesassociated with yeast or fungi comprising admixing a compound orpolypeptide according to claim 49 with a suitable pharmaceuticallyacceptable carrier.
 51. A pharmaceutical composition comprising acompound or polypeptide according to claim 49 together with a suitablepharmaceutically acceptable carrier.
 52. Use of a compound orpolypeptide according to claim 48 or 49 or a pharmaceutical compositionaccording to claim 51 or obtainable by the method of claim 50 for thepreparation of a medicament for treating diseases associated with yeastand fungi.
 53. A method for preventing infection with yeast or fungicomprising administering a composition according to claim 51 orobtainable by the method of claim 50 to a mammal in an effective amountto stimulate the production of protective antibody or protective T-cellresponse.
 54. Use of an antibody capable of specifically binding to atleast one of the polypeptides as defined in claim 37 or to a specificepitope of said polypeptide, for the preparation of a medicament fortreating diseases associated with yeast and fungi.
 55. Use according toany of claims 52 to 54 wherein said disease is associated with yeast orfungi, where the yeast or fungus is chosen from Candida spp.,Aspergillus spp., Microsporum spp., Trichophyton spp., Fusarium spp.,Zygomycetes spp., Botritis, spp., Cladosporium spp., Malassezia spp.,Epidermophyton floccosum, Blastomyces dermatitidis, Coccidioidesimmitis, Histoplasma capsulatum, Paracoccidioides brasiliensis,Cryptococcus neoformans, and Sporothrix schenckii.
 56. Use of a compoundor polypeptide according to claim 48 or 49 or a pharmaceuticalcomposition according to claim 51 or a genetically modified organism asdefined in claim 40 for the preparation of a medicament for modifyingthe endogenic flora of humans and other mammals.
 57. A geneticallymodified mammalian cell or non-human organism in which modificationresults in the overexpression or underexpression of at least one of thenucleic acids as defined in claim 36 or a human homologue thereof or atleast one of the polypeptides as defined in claim 37 or a humanhomologue thereof, which overexpression or underexpression of saidnucleic acid or polypeptide prevents, delays or sensitizes for apoptosisof said genetically modified mammalian cell or in said geneticallymodified non-human organism.
 58. A genetically modified mammalian cellor non-human organism according to claim 57 wherein said modificationcomprises the expression of an antisense molecule to at least one of thenucleic acids as defined in claim 36 or an antisense molecule to amammalian homologue of said nucleic acid.
 59. A method for identifyingcompounds for stimulating or inhibiting apoptosis comprising the use ofat least one of the nucleic acids as defined in claim 36 or a humanhomologue thereof and/or at least one of the polypeptides as defined inclaim 37 or a human homologue thereof and/or a genetically modifiedmammalian cell or non-human organism according to claim 57 or
 58. 60. Acompound identifiable according to the method of claim
 59. 61. Acompound according to claim 60 for use as a medicament.
 62. A method forpreparing a pharmaceutical composition for treating proliferativedisorders or for preventing apoptosis in certain diseases comprisingadmixing a compound according to claim 60 or 61 with a suitablepharmaceutically acceptable carrier.
 63. Use of a compound according toclaim 60 or 61 for the preparation of a medicament for treatingproliferative disorders or for preventing apoptosis in certaindisorders.
 64. Use of a nucleic acid selected from any of the nucleicacids as defined in claim 36 or a human homologue thereof for treatingan/or preventing and/or alleviating proliferative disorders or for theprevention of apoptosis in certain diseases.
 65. Use of a nucleic acidselected from any of the nucleic acids as defined in claim 36 or a humanhomologue thereof for the preparation of a medicament for treatingand/or preventing and/or alleviating proliferative disorders or for theprevention of apoptosis in certain diseases.
 66. Use of an antisensemolecule to at least one of the nucleic acids as defined in claim 36 oran antisense molecule to a mammalian homologue of said nucleic acid fortreating and/or preventing and/or alleviating proliferative disorders orfor preventing apoptosis in certain disorders.
 67. Use of an antisensemolecule to at least one of the nucleic acids as defined in claim 36 oran antisense molecule to a mammalian homologue of said nucleic acid forthe preparation of a medicament for treating and/or preventing and/oralleviating proliferative disorders or for preventing apoptosis incertain disorders.
 68. Use of a polypeptide selected from any of thepolypeptides as defined in claim 37 or a human homologue thereof fortreating and/or preventing and/or alleviating proliferative disorders orfor the prevention of apoptosis in certain diseases.
 69. Apharmaceutical composition for use as a medicament for treatingproliferative disorders or for the prevention of apoptosis in certaindiseases comprising a nucleic acid molecule as defined in claim 36 or ahuman homologue thereof or an antisense molecule to at least one of thenucleic acids as defined in claim 36 or an antisense molecule to amammalian homologue of said nucleic acid or a polypeptide as defined inclaim 37 or a human homologue thereof together with a pharmaceuticallyacceptable carrier diluent or excipient therefor.
 70. A vaccine forimmunizing mammals against proliferative disorders or for preventingapoptosis in certain diseases comprising least one nucleic acid asdefined in claim 36 or a human homologue thereof or at least onepolypeptide as defined in claim 37 or a human analogue thereof in apharmaceutically acceptable carrier.
 71. Use of an antibody capable ofspecifically binding to at least one of the polypeptides as defined inclaim 37 or to a human homologue thereof or to a specific epitope ofsaid polypeptide or said human homologue, for the preparation of amedicament for treating proliferative disorders or for the prevention ofapoptosis in certain diseases.
 72. An expression vector comprising ahuman homologue of a nucleic acid as defined in claim
 36. 73. Anexpression vector according to claim 72 which is an expression vectorwherein said nucleic acid sequence is operably linked to one or morecontrol sequences allowing the expression in prokaryotic and/oreukaryotic host cells.
 74. An expression vector according to claim 72 or73 which comprises an inducible promoter.
 75. An expression vectoraccording to any of claims 72 to 74 which comprises a sequence encodinga reporter molecule.
 76. A host cell transformed, transfected orinfected with the vector of any of claims 72 to
 75. 77. An isolatednucleic acid comprising a human homologue of at least one of the nucleicacids as defined in claim
 36. 78. An antisense molecule comprising anucleic acid sequence capable of selectively hybridising to the nucleicacid molecule of claim
 77. 79. A polypeptide encoded by a nucleic acidof claim 77.